ARGENT Regulated Homodimerization

2006

Bashamboo, A., Taylor, A.H., Samuel, K., Panthier, J.J., Whetton, A.D., and Forrester, L.M. (2006) The survival of differentiating embryonic stem cells is dependent on the SCF-KIT pathway. J Cell Sci 119:3039-3046.
Studies the function of the stem cell factor receptor KIT in embryonic stems cells by introducing an AP20187-inducible receptor into Kit-null cells.
Burnett, S.H., Beus, B.J., Avdiushko, R., Qualls, J., Kaplan, A.M., and Cohen, D.A. (2006) Development of peritoneal adhesions in macrophage depleted mice. J Surg Res 131:296-301.
Develops a mouse model to study the induction and repair of peritoneal adhesions based on the finding that such adhesions develop upon AP20187-mediated depletion of macrophages in the previously described "Mafia" mice (Burnett et al., 2004).
Clackson, T. (2006) Dissecting the Functions of Proteins and Pathways using Chemically Induced Dimerization. Chem Biol Drug Des 67:440-442.
Briefly reviews the principles and uses of the ARGENT and RPD systems for chemically-controlled dimerization.
Fooksman, D.R., Gronvall, G.K., Tang, Q., and Edidin, M. (2006) Clustering class I MHC modulates sensitivity of T cell recognition. J Immunol 176:6673-6680.
Uses AP20187-mediated dimerization to show that clustering of class I MHC molecules on the surface of B lymphoblasts enhances their recognition by T cells.
Fukada, M., Fujikawa, A., Chow, J.P., Ikematsu, S., Sakuma, S., and Noda, M. (2006) Protein tyrosine phosphatase receptor type Z is inactivated by ligand-induced oligomerization. FEBS Lett 580:4051-4056.
Uses AP20187 to demonstrate that a protein tyrosine phosphatase receptor is inactivated by oligomerization.
Hirate, Y., and Okamoto, H. (2006) Canopy1, a Novel Regulator of FGF Signaling around the Midbrain-Hindbrain Boundary in Zebrafish. Curr Biol 16:421-427.
Uses AP20187-induced dimerization of FGFR1 to demonstrate that expression of Canopy1 is essential for normal FGF signaling in zebrafish embryos. The inducible FGFR1 gene was injected as mRNA into a specific area of the brain and AP20187 was added directly to the embryos.
Marciniak, S.J., Garcia-Bonilla, L., Hu, J., Harding, H.P., and Ron, D. (2006) Activation-dependent substrate recruitment by the eukaryotic translation initiation factor 2 kinase PERK. J Cell Biol 172:201-209.
Uses AP20187-mediated oligomerization of PERK to study its mechanism of activation and downstream signalling.
Nagasawa, Y., Wood, B.L., Wang, L., Lintmaer, I., Guo, W., Papayannopoulou, T., Harkey, M.A., Nourigat, C., and Blau, C.A. (2006) Anatomical compartments modify the response of human hematopoietic cells to a mitogenic signal. Stem Cells 24:908-917.
Uses AP20187-mediated oligomerization of mpl to drive expansion of human cord blood CD34+ cells transplanted into mice. It is found that response of the engineered cells to the mitogenic signal is influenced by the anatomical compartment in which they reside.
Pelletier, L., Guillaumot, P., Freche, B., Luquain, C., Christiansen, D., Brugiere, S., Garin, J., and Manie, S.N. (2006) Gamma-secretase-dependent proteolysis of CD44 promotes neoplastic transformation of rat fibroblastic cells. Cancer Res 66:3681-3687.
Uses AP20187-controlled homodimerization of Ret to show that Ret-driven neoplastic transformation correlates with increased expression and subsequent cleavage of CD44.
Pinsky, M.S., Song, W., Dong, Z., Warner, K., Zeitlin, B., Karl, E., Hall, D.E., and Nor, J.E. (2006) Activation of iCaspase-9 in neovessels inhibits oral tumor progression. J Dent Res 85:436-441.
Uses AP20187-mediated dimerization to show that activation of caspase-9 in neovascular endothelial cells is sufficient to inhibit growth of oral tumors xenografted into mice.
Schwertfeger, K.L., Xian, W., Kaplan, A.M., Burnett, S.H., Cohen, D.A., and Rosen, J.M. (2006) A critical role for the inflammatory response in a mouse model of preneoplastic progression. Cancer Res 66:5676-5685.
Describes the use of transgenic mice that express an AP20187-inducible fibroblast growth factor receptor-1 (iFGFR1) to examine role of the microenvironment in early stages of tumorigenesis. These mice were also crossed with "Mafia" transgenic mice that contain an AP20187-inducible apoptotic switch expressed in macrophages to study the effect that macrophage depletion has on iFGFR1-mediated phenotypes.
Sun, M., Ames, K.T., Suzuki, I., and Fink, P.J. (2006) The cytoplasmic domain of Fas ligand costimulates TCR signals. J Immunol 177:1481-1491.
Uses FK1012 and AP1510 to demonstrate that oligomerization of the FasL cytoplasmic domain is sufficient for FasL-mediated costimulation of T cells.
Xiao, H., Wang, L.L., Shu, C.L., Yu, M., Li, S., Shen, B.F., and Li, Y. (2006) Establishment of a Cell Model Based on FKBP12 Dimerization for Screening of FK506-like Neurotrophic Small Molecular Compounds. J Biomol Screen.
Uses the AP20187-mediated homodimerization system as the basis of a screen for novel FK506-like small molecules. Compounds are screened for the ability to block apoptosis caused by forced dimerization of mBax.
Zhan, L., Xiang, B., and Muthuswamy, S.K. (2006) Controlled activation of ErbB1/ErbB2 heterodimers promote invasion of three-dimensional organized epithelia in an ErbB1-dependent manner: implications for progression of ErbB2-overexpressing tumors. Cancer Res 66:5201-5208.
Uses AP1510-mediated homodimerization and AP21967-mediated heterodimerization to compare the tumor-promoting activities of ErbB2 homodimers and ErbB1-ErbB2 heterodimers.

2005

Abell, A.N., and Johnson, G.L. (2005) MEKK4 is an effector of the embryonic TRAF4 for JNK activation. J Biol Chem 280:35793-35796.
Uses AP20187 to show that oligomerization of MEKK4 is sufficient to activate JNK.
Blau, C.A., Barbas, C.F., 3rd, Bomhoff, A.L., Neades, R., Yan, J., Navas, P.A., and Peterson, K.R. (2005) {gamma}-Globin gene expression in chemical inducer of dimerization (CID)-dependent multipotential cells established from human {beta}-globin locus yeast artificial chromosome ({beta}-YAC) transgenic mice. J Biol Chem 280:36642-36647.
Developed cells that can be used to screen for inducers of gamma-globulin expression by using an AP20187-inducible mpl construct to drive proliferation of bone marrow cells derived from beta-YAC transgenic mice.
Carlotti, F., Zaldumbide, A., Martin, P., Boulukos, K.E., Hoeben, R.C., and Pognonec, P. (2005) Development of an inducible suicide gene system based on human caspase 8. Cancer Gene Ther 12:627-639.
Uses AP20187 to demonstrate that control of caspase 8 oligomerization allows for tightly regulated induction of apoptosis.
Cheng, J., Yu, L., Zhang, D., Huang, Q., Spencer, D., and Su, B. (2005) Dimerization through the catalytic domain is essential for MEKK2 activation. J Biol Chem 280:13477-13482.
Uses AP20187 to demonstrate that oligomerization of MEKK2 leads to its activation.
Emery, D.W., Tubb, J., Nishino, Y., Nishino, T., Otto, K.G., Stamatoyannopoulos, G., and Blau, C.A. (2005) Selection with a regulated cell growth switch increases the likelihood of expression for a linked gamma-globin gene. Blood Cells Mol Dis 34:235-247.
In vivo expansion of hematopoietic stem cells using the AP20187-inducible Mpl growth switch increases expression of a linked gamma-globin gene.
Freche, B., Guillaumot, P., Charmetant, J., Pelletier, L., Luquain, C., Christiansen, D., Billaud, M., and Manie, S.N. (2005) Inducible dimerization of RET reveals a specific AKT deregulation in oncogenic signaling. J Biol Chem 280:36584-36591.
Uses AP20187-mediated dimerization to study downstream effects of activating the receptor tyrosine kinase RET.
Gazdoiu, S., Yamoah, K., Wu, K., Escalante, C.R., Tappin, I., Bermudez, V., Aggarwal, A.K., Hurwitz, J., and Pan, Z.Q. (2005) Proximity-induced activation of human Cdc34 through heterologous dimerization. Proc Natl Acad Sci U S A 102:15053-15058.
Demonstrates that AP20187-mediated dimerization of Cdc34 (overexpressed and purified from insect cells) is sufficient to activate its catalytic activity.
Gouzi, J.Y., Moog-Lutz, C., Vigny, M., and Brunet-de Carvalho, N. (2005) Role of the subcellular localization of ALK tyrosine kinase domain in neuronal differentiation of PC12 cells. J Cell Sci 118:5811-5823.
Uses AP20187 to control the dimerization and activation of the membrane tyrosine kinase ALK.
Hanks, B.A., Jiang, J., Singh, R.A., Song, W., Barry, M., Huls, M.H., Slawin, K.M., and Spencer, D.M. (2005) Re-engineered CD40 receptor enables potent pharmacological activation of dendritic-cell cancer vaccines in vivo. Nat Med 11:130-137.
Dendritic cell activation mediated by an AP20187-inducible CD40 receptor results in more potent T-cell effector responses in mice and has the potential to lead to more potent human cancer vaccines.
Harding, H.P., Zhang, Y., Khersonsky, S., Marciniak, S., Scheuner, D., Kaufman, R.J., Javitt, N., Chang, Y.T., and Ron, D. (2005) Bioactive small molecules reveal antagonism between the integrated stress response and sterol-regulated gene expression. Cell Metab 2:361-371.
Uses AP20187-inducible PERK to examine the role that phosphorylation of its substrate, eIF2alpha, plays in the inhibition of sterol regulatory element binding protein target genes.
Karpova, A.Y., Tervo, D.G., Gray, N.W., and Svoboda, K. (2005) Rapid and reversible chemical inactivation of synaptic transmission in genetically targeted neurons. Neuron 48:727-735.
Describes the development of dimerization-based switches, called MISTs, for reversible inactivation of neurotransmitter release. The switches are based on sequestration of proteins essential to synaptic vesicle fusion, using AP20187-mediated homodimerization or AP21967-mediated heterodimerization. Switches functioned in cultured neurons, brain slices and (for AP20187 injected into the brain) in transgenic mice expressing MISTs selectively in Purkinje neurons.
Kobinger, G.P., Deng, S., Louboutin, J.P., Vatamaniuk, M., Rivera, V.M., Lian, M.M., Markmann, J.F., Clackson, T., Raper, S.E., Matschinsky, F., and Wilson, J.M. (2005) Pharmacologically regulated regeneration of functional human pancreatic islets. Mol Ther 11:105-111.
A method was developed to stimulate proliferation of insulin secreting beta-cells in vitro and in vivo using an AP20187-inducible Epo receptor.
Larrivee, B., Niessen, K., Pollet, I., Corbel, S.Y., Long, M., Rossi, F.M., Olive, P.L., and Karsan, A. (2005) Minimal contribution of marrow-derived endothelial precursors to tumor vasculature. J Immunol 175:2890-2899.
Uses an AP20187-inducible VEGF receptor 2 to demonstrate that the VEGFR-2 pathway is not sufficient for the recruitment and/or expansion of endothelial progenitor cells in mice.
Larrivee, B., Pollet, I., and Karsan, A. (2005) Activation of vascular endothelial growth factor receptor-2 in bone marrow leads to accumulation of myeloid cells: role of granulocyte-macrophage colony-stimulating factor. J Immunol 175:3015-3024.
Uses an AP20187-inducible VEGF receptor 2 to demonstrate that the VEGFR-2 pathway induces expansion of myeloid cells in mice.
Lupo, G., Liu, Y., Qiu, R., Chandraratna, R.A., Barsacchi, G., He, R.Q., and Harris, W.A. (2005) Dorsoventral patterning of the Xenopus eye: a collaboration of Retinoid, Hedgehog and FGF receptor signaling. Development 132:1737-1748.
Uses AP20187-inducible FGF receptor 1 to explore the role of the FGFR1 signaling pathway in dorsoventral patterning of the Xenopus eye.
Nikitina, E.Y., Desai, S.A., Zhao, X., Song, W., Luo, A.Z., Gangula, R.D., Slawin, K.M., and Spencer, D.M. (2005) Versatile prostate cancer treatment with inducible caspase and interleukin-12. Cancer Res 65:4309-4319.
Further explored the use of AP20187-mediated oligomerization of caspase-1 to induce tumor cell apoptosis in a mouse prostate cancer model.
Pajvani, U.B., Trujillo, M.E., Combs, T.P., Iyengar, P., Jelicks, L., Roth, K.A., Kitsis, R.N., and Scherer, P.E. (2005) Fat apoptosis through targeted activation of caspase 8: a new mouse model of inducible and reversible lipoatrophy. Nat Med 11:797-803.
Describes the generation of transgenic mice expressing an FKBP-caspase 8 fusion protein that allows selective elimination of adipocytes by administration of AP20187.
Reginato, M.J., Mills, K.R., Becker, E.B., Lynch, D.K., Bonni, A., Muthuswamy, S.K., and Brugge, J.S. (2005) Bim regulation of lumen formation in cultured mammary epithelial acini is targeted by oncogenes. Mol Cell Biol 25:4591-4601.
Uses AP1510-mediated oligomerization of ErbB2 to study the role of Bim in the regulation of lumen formation.
Siatskas, C., Underwood, J., Ramezani, A., Hawley, R.G., and Medin, J.A. (2005) Specific pharmacological dimerization of KDR in lentivirally transduced human hematopoietic cells activates antiapoptotic and proliferative mechanisms. Faseb J.
Uses AP20187 to study the function of a lentivirally transduced KDR/flk-1/VEGFR-2 chimeric receptor using functional, biochemical and microarray analyses.
Song, G.J., and Hinkle, P.M. (2005) Regulated Dimerization of the Thyrotropin-Releasing Hormone Receptor Affects Receptor Trafficking but Not Signaling. Mol Endocrinol.
Uses AP20187 to examine the effect that dimerization has on the trafficking of the thyrotropin-releasing hormone receptor, a G protein-coupled receptor. An excess of the monomeric ligand AP21998 is shown to block the effect.
Song, W., Sun, Q., Dong, Z., Spencer, D.M., Nunez, G., and Nor, J.E. (2005) Antiangiogenic gene therapy: disruption of neovascular networks mediated by inducible caspase-9 delivered with a transcriptionally targeted adenoviral vector. Gene Ther 12:320-329.
Delivery of an AP20187-inducible caspase-9 to endothelial cells of mice using a transcriptionally targeted adenoviral vector results in disruption of neovascularization.
Storez, H., Scott, M.G., Issafras, H., Burtey, A., Benmerah, A., Muntaner, O., Piolot, T., Tramier, M., Coppey-Moisan, M., Bouvier, M., Labbe-Jullie, C., and Marullo, S. (2005) Homo- and hetero-oligomerization of beta-arrestins in living cells. J Biol Chem 280:40210-40215.
Describes the use of AP20187-mediated homodimerization of GFP-labelled beta arrestin 2 to demonstrate that the pre-oligomerized protein has unchanged intracellular translocation kinetics and behavior to the undimerized protein.
Straathof, K.C., Pule, M.A., Yotnda, P., Dotti, G., Vanin, E.F., Brenner, M.K., Heslop, H.E., Spencer, D.M., and Rooney, C.M. (2005) An inducible caspase 9 safety switch for T-cell therapy. Blood 105:4247-4254.
Demonstrates that an AP20187-inducible caspase-9 can lead to the selective elimination of 99% of transduced T cells in vivo, suggesting promise as a safety switch for human T-cell therapies.
Trujillo, M.E., Pajvani, U.B., and Scherer, P.E. (2005) Apoptosis Through Targeted Activation of Caspase8 ("ATTAC-mice"): Novel Mouse Models of Inducible and Reversible Tissue Ablation. Cell Cycle 4:1141-1145.
Reviews the use of the AP21087 system to develop mouse models in which adipocytes and other specific cell types can be inducibly eliminated via dimerization of caspase-8.
Van Stry, M., Kazlauskas, A., Schreiber, S.L., and Symes, K. (2005) Distinct effectors of platelet-derived growth factor receptor-alpha signaling are required for cell survival during embryogenesis. Proc Natl Acad Sci U S A 102:8233-8238.
Uses AP1510-mediate dimerization of PDGFR-alpha to study the effectors of its activity in Xenopus embryos.

2004

Berger, C., Blau, C.A., Huang, M.L., Iuliucci, J.D., Dalgarno, D.C., Gaschet, J., Heimfeld, S., Clackson, T., and Riddell, S.R. (2004) Pharmacologically regulated Fas-mediated death of adoptively transferred T cells in a nonhuman primate model. Blood 103:1261-1269.
Demonstrates that the AP1903-inducible Fas-based suicide switch can be employed to regulate the survival of adoptively transferred T cells in non-human primates.
Brouckaert, G., Kalai, M., Krysko, D.V., Saelens, X., Vercammen, D., Ndlovu, Haegeman, G., D'Herde, K., and Vandenabeele, P. (2004) Phagocytosis of necrotic cells by macrophages is phosphatidylserine dependent and does not induce inflammatory cytokine production. Mol Biol Cell 15:1089-1100.
Uses AP1510-mediated oligomerization of an FKBP-FADD fusion protein to induce necrotic cell death.
Burnett, S.H., Kershen, E.J., Zhang, J., Zeng, L., Straley, S.C., Kaplan, A.M., and Cohen, D.A. (2004) Conditional macrophage ablation in transgenic mice expressing a Fas-based suicide gene. J Leukoc Biol 75:612-623.
Describes the generation of transgenic mice that express an AP20187-inducible Fas death switch selectively in macrophages and dendritic cells. These mice allow the effects of the selective depletion of macrophages and dendritic cells to be studied in vivo.
Cotugno, G., Pollock, R., Formisano, P., Linher, K., Beguinot, F., and Auricchio, A. (2004) Pharmacological regulation of the insulin receptor signaling pathway mimics insulin action in cells transduced with viral vectors. Hum Gene Ther 15:1101-1108.
A system for pharmacologic regulation of the insulin signaling pathway is described based on AP20187-mediated oligomerization of the insulin receptor.
Deng, J., Lu, P.D., Zhang, Y., Scheuner, D., Kaufman, R.J., Sonenberg, N., Harding, H.P., and Ron, D. (2004) Translational repression mediates activation of nuclear factor kappa B by phosphorylated translation initiation factor 2. Mol Cell Biol 24:10161-10168.
Uses AP20187-mediated activation of PERK to establish a role for eIF2-alpha phosphorylation in NF-kappaB activation.
Iwamoto, K., Miyamoto, T., Sawatani, Y., Hosogane, N., Hamaguchi, I., Takami, M., Nomiyama, K., Takagi, K., and Suda, T. (2004) Dimer formation of receptor activator of nuclear factor kappaB induces incomplete osteoclast formation. Biochem Biophys Res Commun 325:229-234.
AP20187-mediated dimerization of the RANK receptor induces a subset of the activities induced by the RANK ligand induced trimerization.
Karlsson, T., Henriksson, R., and Hedman, H. (2004) Induction of apoptosis in resistant glioma cells by synthetic caspase-activation. J Neurooncol 66:71-79.
Uses AP20187-inducible FADD and caspases 3 and 8 to delineate the molecular basis of apoptosis resistance of several glioma cell lines.
Lu, P.D., Jousse, C., Marciniak, S.J., Zhang, Y., Novoa, I., Scheuner, D., Kaufman, R.J., Ron, D., and Harding, H.P. (2004) Cytoprotection by pre-emptive conditional phosphorylation of translation initiation factor 2. Embo J 23:169-179.
Uses AP20187-mediated oligomerization of PERK to study the activity of PERK independent of upstream stress signaling.
Mikolajczyk, J., Scott, F.L., Krajewski, S., Sutherlin, D.P., and Salvesen, G.S. (2004) Activation and substrate specificity of caspase-14. Biochemistry 43:10560-10569.
Studies the role that dimerization has in controlling caspase 14 activity using the AP20187-mediated homodimerization system.
Nam, Y.J., Mani, K., Ashton, A.W., Peng, C.F., Krishnamurthy, B., Hayakawa, Y., Lee, P., Korsmeyer, S.J., and Kitsis, R.N. (2004) Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions. Mol Cell 15:901-912.
Demonstrates that AP20187-mediated dimerization of ARC decreases ARC's ability to inhibit apoptosis.
Richard, R.E., De Claro, R.A., Yan, J., Chien, S., Von Recum, H., Morris, J., Kiem, H.P., Dalgarno, D.C., Heimfeld, S., Clackson, T., Andrews, R., and Blau, C.A. (2004) Differences in F36VMpl-based in vivo selection among large animal models. Mol Ther 10:730-740.
AP20187-mediated oligomerization of mpl leads to only a modest expansion of hematopoietic cells in baboons in contrast to what was seen previously in mice.
Richard, R.E., Weinreich, M., Chang, K.H., Ieremia, J., Stevenson, M.M., and Blau, C.A. (2004) Modulating erythrocyte chimerism in a mouse model of pyruvate kinase deficiency. Blood 103:4432-4439.
AP20187-mediated oligomerization of mpl is used to show that expansion of donor red blood cells can improve inherited anemia and reduce abnormal erythropoiesis in mice.
Seton-Rogers, S.E., Lu, Y., Hines, L.M., Koundinya, M., LaBaer, J., Muthuswamy, S.K., and Brugge, J.S. (2004) Cooperation of the ErbB2 receptor and transforming growth factor beta in induction of migration and invasion in mammary epithelial cells. Proc Natl Acad Sci U S A101:1257-1262.
Describes the results of a screen for genes that induce migration of cells already activated by AP1510-mediated dimerization of ErbB2.
Strasser, V., Fasching, D., Hauser, C., Mayer, H., Bock, H.H., Hiesberger, T., Herz, J., Weeber, E.J., Sweatt, J.D., Pramatarova, A., Howell, B., Schneider, W.J., and Nimpf, J. (2004) Receptor Clustering Is Involved in Reelin Signaling. Mol Cell Biol 24:1378-1386.
AP20187-mediated dimerization of Dab1 is sufficient to induce its phosphorylation.
Tognon, C.E., Mackereth, C.D., Somasiri, A.M., McIntosh, L.P., and Sorensen, P.H. (2004) Mutations in the SAM domain of the ETV6-NTRK3 chimeric tyrosine kinase block polymerization and transformation activity. Mol Cell Biol 24:4636-4650.
Uses AP20187 to show that oligomerization of NTRK3 is not sufficient to transform cells.
Vanden Berghe, T., Van Loo, G., Saelens, X., Van Gurp, M., Brouckaert, G., Kalai, M., Declercq, W., and Vandenabeele, P. (2004) Differential signaling to apoptotic and necrotic cell death by Fas-associated death domain protein FADD. J Biol Chem 279:7925-7933.
Uses AP1510-mediated oligomerization to study the role of FADD in apoptotic and necrotic signaling pathways.
Wang, S., Ai, X., Freeman, S.D., Pownall, M.E., Lu, Q., Kessler, D.S., and Emerson, C.P., Jr. (2004) QSulf1, a heparan sulfate 6-O-endosulfatase, inhibits fibroblast growth factor signaling in mesoderm induction and angiogenesis. Proc Natl Acad Sci U S A 101:4833-4838.
Uses AP20187-mediated oligomerization of FGFR1 to study its function in xenopus embryos.
Zhao, S., Weinreich, M.A., Ihara, K., Richard, R.E., and Blau, C.A. (2004) In vivo selection of genetically modified erythroid cells using a jak2-based cell growth switch. Mol Ther 10:456-468.
Demonstrates that AP20187-mediated oligomerization of Jak2 acts as an erythroid-specific cell growth switch upon transduction of murine marrow cells in mice.

2003

Arias-Salgado, E.G., Lizano, S., Sarkar, S., Brugge, J.S., Ginsberg, M.H., and Shattil, S.J. (2003) Src kinase activation by direct interaction with the integrin {beta} cytoplasmic domain. Proc Natl Acad Sci USA 100:13298-13302.
Uses AP1510 to show that Src can be activated via clustering of beta3 integrins.
Arya, M., Lopez, J.A., Romo, G.M., Cruz, M.A., Kasirer-Friede, A., Shattil, S.J., and Anvari, B. (2003) Glycoprotein Ib-IX-mediated activation of integrin alpha(IIb)beta(3): effects of receptor clustering and von Willebrand factor adhesion. J Thromb Haemost 1:1150-7.
AP20187-mediated dimerization of glycoprotein Ib-IX increases the strength of its interaction with the von Willebrand factor.
Berger, C., Blau, C.A., Clackson, T., Riddell, S.R., and Heimfeld, S. (2003) CD28 costimulation and immunoaffinity-based selection efficiently generate primary gene-modified T cells for adoptive immunotherapy. Blood 101:476-84.
Describes the development of a strategy to generate large numbers of T cells transduced with a retroviral vector encoding a dimerizer-activated Fas-based suicide gene, and characterization of the functionality of the resulting cells.
Buensuceso, C., de Virgilio, M., and Shattil, S.J. (2003) Detection of integrin alpha IIbbeta 3 clustering in living cells. J Biol Chem 278:15217-24.
Uses AP1510 to study the clustering of integrin alphaIIb-beta3 induced by intracellular events.
Carlson, J.C., Kanter, A., Thuduppathy, G.R., Cody, V., Pineda, P.E., McIvor, R.S., and Wagner, C.R. (2003) Designing protein dimerizers: the importance of ligand conformational equilibria. J Am Chem Soc 125:1501-7.
Describes theoretical, biophysical, and structural analysis of a methotrexate homodimerizer, including a determination of the thermodynamics of dimerization and the crystal structure of the complex between the dimerizer and two copies of E. coli dihydrofolate reductase (DHFR).
Chang, D.W., Ditsworth, D., Liu, H., Srinivasula, S.M., Alnemri, E.S., and Yang, X. (2003) Oligomerization is a general mechanism for the activation of apoptosis initiator and inflammatory procaspases. J Biol Chem 278:16466-9.
Uses AP20187 to demonstrate that both initiator (e.g. -9 and -10) and inflammatory (e.g. -11) caspases are activated by oligomerization.
Chang, D.W., Xing, Z., Capacio, V.L., Peter, M.E., and Yang, X. (2003) Interdimer processing mechanism of procaspase-8 activation. EMBO J 22:4132-42.
Uses AP20187 to reveal key dimerization-based steps that lead to the activation of procaspase-8.
Chang, D.W., and Yang, X. (2003) Activation of procaspases by FK506 binding protein-mediated oligomerization. Sci STKE 2003:PL1.
Provides protocols for achieving dimerizer-inducible apoptosis of mammalian cells using FKBP-caspase-8 fusion proteins and AP20187.
Dowds, T.A., Masumoto, J., Chen, F.F., Ogura, Y., Inohara, N., and Nunez, G. (2003) Regulation of cryopyrin/Pypaf1 signaling by pyrin, the familial Mediterranean fever gene product. Biochem Biophys Res Commun 302:575-80.
Studies using the homodimerizer AP1510 reveal interactions between cryopyrin and ASC that lead to apoptosis.
Freeman, K.W., Gangula, R.D., Welm, B.E., Ozen, M., Foster, B.A., Rosen, J.M., Ittmann, M., Greenberg, N.M., and Spencer, D.M. (2003) Conditional activation of fibroblast growth factor receptor (FGFR) 1, but not FGFR2, in prostate cancer cells leads to increased osteopontin induction, extracellular signal-regulated kinase activation, and in vivo proliferation. Cancer Res 63:6237-43.
Uses an AP20187-inducible FGFR1 to demonstrate its role in signalling and its ability to promote growth of prostate tumor cells in vivo.
Freeman, K.W., Welm, B.E., Gangula, R.D., Rosen, J.M., Ittmann, M., Greenberg, N.M., and Spencer, D.M. (2003) Inducible prostate intraepithelial neoplasia with reversible hyperplasia in conditional FGFR1-expressing mice. Cancer Res 63:8256-63.
Uses transgenic mice containing an AP20187-inducible FGFR1 to show that the development and progression of key pathologic changes seen in early-stage prostate cancer are directly dependent on FGFR1 activation, and that the resultant early pathologies are completely reversible.
Godbey, W.T., and Atala, A. (2003) Directed apoptosis in Cox-2-overexpressing cancer cells through expression-targeted gene delivery. Gene Ther 10:1519-27.
Uses an AP20187-inducible caspase-3 (or -9), driven by a Cox-2-inducible promoter, to selectively induce apoptosis in tumor cells that overexpress Cox-2, including cells that are typically resistant to apoptosis.
Junker, K., Koehl, U., Zimmerman, S., Stein, S., Schwabe, D., Klingebiel, T., and Grez, M. (2003) Kinetics of cell death in T lymphocytes genetically modified with two novel suicide fusion genes. Gene Ther 10:1189-97.
Demonstrates the efficient and rapid induction of apoptosis in T cells upon AP20187-mediated oligomerization of the death effector domain of FADD.
Kazansky, A.V., Spencer, D.M., and Greenberg, N.M. (2003) Activation of signal transducer and activator of transcription 5 is required for progression of autochthonous prostate cancer: evidence from the transgenic adenocarcinoma of the mouse prostate system. Cancer Res 63:8757-62.
Describes an AP20187-inducible version of a naturally occuring dominant-negative isoform of STAT5B and demonstrates its ability to block the invasive potential of prostate cells.
Larrivee, B., Lane, D.R., Pollet, I., Olive, P.L., Humphries, R.K., and Karsan, A. (2003) Vascular endothelial growth factor receptor-2 induces survival of hematopoietic progenitor cells. J Biol Chem 278:22006-13.
Uses AP20187 to dimerize the intracellular domain of the VEGF receptor-2 to demonstrate its role in maintaining survival of hematopoietic progenitor cells.
Lee, L.A., Van Hoewyk, D., and Orr-Weaver, T.L. (2003) The Drosophila cell cycle kinase PAN GU forms an active complex with PLUTONIUM and GNU to regulate embryonic divisions. J Genes Dev 17:2979-91.
Uses an AP20187-inducible form of the Drosophila protein PLU to study the mechanism of activation of an associated kinase.
Martin, M.E., Milne, T.A., Bloyer, S., Galoian, K., Shen, W., Gibbs, D., Brock, H.W., Slany, R., and Hess, J.L. (2003) Dimerization of MLL fusion proteins immortalizes hematopoietic cells. Cancer Cell 4:197-207.
Demonstrates that AP20187-mediated dimerization of an MLL fusion protein converts it into a transcriptional transactivator and is sufficient to immortalize cells in a reversible manner
Masumoto, J., Dowds, T.A., Schaner, P., Chen, F.F., Ogura, Y., Li, M., Zhu, L., Katsuyama, T., Sagara, J., Taniguchi, S., Gumucio, D.L., Nunez, G., and Inohara, N. (2003) ASC is an activating adaptor for NF-kappa B and caspase-8-dependent apoptosis. Biochem Biophys Res Commun 303:69-73.
Demonstrates that AP1510-induced oligomerization of the caspase recruitment domain of either ASC or Ipaf induces apoptosis and NF-kappaB activation.
Masumoto, J., Zhou, W., Chen, F.F., Su, F., Kuwada, J.Y., Hidaka, E., Katsuyama, T., Sagara, J., Taniguchi, S., Ngo-Hazelett, P., Postlethwait, J.H., Nunez, G., and Inohara, N. (2003) Caspy, a zebrafish caspase, activated by ASC oligomerization is required for pharyngeal arch development. J Biol Chem 278:4268-76.
AP1510-mediated oligomerization of ASC is sufficient to activate the Zebrafish caspase, Caspy.
Moskowitz, H.S., Heuser, J., McGraw, T.E., and Ryan, T.A. (2003) Targeted chemical disruption of clathrin function in living cells. Mol Biol Cell 14:4437-47.
Uses FK1012-mediated homodimerization of clathrin to interfere with its normal assembly-disassembly cycle, allowing clathrin's role in membrane trafficking to be explored.
Muromoto, R., Yamamoto, T., Yumioka, T., Sekine, Y., Sugiyama, K., Shimoda, K., Oritani, K., and Matsuda, T. (2003) Daxx enhances Fas-mediated apoptosis in a murine pro-B cell line, BAF3. FEBS Lett 540:223-8.
Uses an AP20187-inducible Fas death switch to study the roles of JNK activation and the Fas-associated protein Daxx in inducing apoptosis in B cells.
Perlingeiro, R.C., Kyba, M., Bodie, S., and Daley, G.Q. (2003) A role for thrombopoietin in hemangioblast development. Stem Cells 21:272-80.
AP20187-mediated oligomerization of the c-Mpl signaling domain is shown to enhance hemangioblast formation.
Pownall, M.E., Welm, B.E., Freeman, K.W., Spencer, D.M., Rosen, J.M., and Isaacs, H.V. (2003) An inducible system for the study of FGF signalling in early amphibian development. Dev Biol 256:90-100.
Uses an AP20187-inducible FGF-receptor to study its role during Xenopus laevis development.
Reginato, M.J., Mills, K.R., Paulus, J.K., Lynch, D.K., Sgroi, D.C., Debnath, J., Muthuswamy, S.K., and Brugge, J.S. (2003) Integrins and EGFR coordinately regulate the pro-apoptotic protein Bim to prevent anoikis. Nat Cell Biol 5:733-40.
AP1510-mediated oligomerization of ErbB2 is shown to inhibit expression of the pro-apoptotic protein Bim and confer resistance to anoikis.
Richard, R.E., and Blau, C.A. (2003) Small-molecule-directed mpl signaling can complement growth factors to selectively expand genetically modified cord blood cells. Stem Cells 21:71-8.
AP20187-mediated oligomerization of mpl allows selective expansion of human cord blood cells when the dimerizer-induced signal is supplemented by additional signals provided by growth factors.
So, C.W., Lin, M., Ayton, P.M., Chen, E.H., and Cleary, M.L. (2003) Dimerization contributes to oncogenic activation of MLL chimeras in acute leukemias. Cancer Cell 4:99-110.
Demonstrates that AP21967-mediated dimerization of MLL is sufficient to enhance the self-renewal of hematopoietic progenitors and provides support for an oligomerization-dependent mechanism for oncogenic conversion of MLL.
Tang, E.D., Inohara, N., Wang, C.Y., Nunez, G., and Guan, K.L. (2003) Roles for homotypic interactions and transautophosphorylation in IkappaB kinase (IKKbeta) activation. J Biol Chem 278:38566-70.
Uses AP1510-mediated oligomerization to help establish that the activation of IKKbeta depends on its ability to homodimerize.
Wencker, D., Chandra, M., Nguyen, K., Miao, W., Garantziotis, S., Factor, S.M., Shirani, J., Armstrong, R.C., and Kitsis, R.N. (2003) A mechanistic role for cardiac myocyte apoptosis in heart failure. J Clin Invest 111:1497-504.
Transgenic mice expressing an FK1012-inducible caspase-8 gene in the myocardium are used to demonstrate, for the first time, that cardiac myocyte apoptosis plays a role in heart failure.

2002

Boccaccio, C., Ando, M., and Comoglio, P.M. (2002) A differentiation switch for genetically modified hepatocytes. Faseb J 16:120-2.
AP1510-mediated oligomerization of the signaling domain of the hepatocyte growth factor receptor induces differentiation, but not proliferation, of hepatocyte progenitor cells.
Chadee, D.N., Yuasa, T., and Kyriakis, J.M. (2002) Direct activation of mitogen-activated protein kinase kinase kinase MEKK1 by the Ste20p homologue GCK and the adapter protein TRAF2. Mol Cell Biol 22:737-49.
AP1510-mediated oligomerization of MEKK1 is sufficient to activate SAPK/JNK.
Chang, D.W., Xing, Z., Pan, Y., Algeciras-Schimnich, A., Barnhart, B.C., Yaish-Ohad, S., Peter, M.E., and Yang, X. (2002) c-FLIP(L) is a dual function regulator for caspase-8 activation and CD95-mediated apoptosis. Embo J 21:3704-3714.
Uses rapamcyin-mediated heterodimerization and AP20187-mediated homodimerization to explore the role of c-FLIP-L in the CD95-mediated apoptotic signaling pathway
Debnath, J., Mills, K.R., Collins, N.L., Reginato, M.J., Muthuswamy, S.K., and Brugge, J.S. (2002) The role of apoptosis in creating and maintaining luminal space within normal and oncogene-expressing mammary acini. Cell 111:29-40.
AP1510-mediated oligomerization of ErbB2 in epithelial cells triggers filling of the luminal space of mammary acini.
Feng, H., Zeng, Y., Graner, M.W., and Katsanis, E. (2002) Stressed apoptotic tumor cells stimulate dendritic cells and induce specific cytotoxic T cells. Blood 100:4108-15.
Uses AP20187-mediated oligomerization of Fas to induce apoptosis to explore how the immune system distinguishes stressed from nonstressed apoptotic tumor cells.
Kasirer-Friede, A., Ware, J., Leng, L., Marchese, P., Ruggeri, Z.M., and Shattil, S.J. (2002) Lateral clustering of platelet GP Ib-IX complexes leads to up- regulation of the adhesive function of Integrin alphaIIbbeta3. J Biol Chem 277:11949-56.
Uses AP20187-mediated oligomerization of GP Ib-IX to show that clustering of this receptor mediates its adhesive and signaling functions.
Kuma, A., Mizushima, N., Ishihara, N., and Ohsumi, Y. (2002) Formation of the ~350 kD Apg12-Apg5-Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast. J Biol Chem 277:18619-25.
Uses AP20187 to show that oligomerization of Apg16 is essential for its role in autophagy in yeast.
Li, Z.Y., Otto, K., Richard, R.E., Ni, S., Kirillova, I., Fausto, N., Blau, C.A., and Lieber, A. (2002) Dimerizer-induced proliferation of genetically modified hepatocytes. Mol Ther 5:420-6.
AP20187-mediated oligomerization of gp130 induces proliferation of hepatocytes in vitro and in vivo.
Mallet, V.O., Mitchell, C., Guidotti, J.E., Jaffray, P., Fabre, M., Spencer, D., Arnoult, D., Kahn, A., and Gilgenkrantz, H. (2002) Conditional cell ablation by tight control of caspase-3 dimerization in transgenic mice. Nat Biotechnol 20:1234-9.
Describes a system for creating animal models of degenerative disease. Transgenic mice are generated which express an FKBP-caspase-3 gene in the liver. Systemic administration of AP20187 induces a dose-dependent, specific ablation of hepatocytes.
Neff, T., Horn, P.A., Valli, V.E., Gown, A.M., Wardwell, S., Wood, B.L., von Kalle, C., Schmidt, M., Peterson, L.J., Morris, J.C., Richard, R.E., Clackson, T., Kiem, H.P., and Blau, C.A. (2002) Pharmacologically regulated in vivo selection in a large animal. Blood 100:2026-31.
Introduction of a mpl-FKBP gene into autologous CD34-positive cells allows reversible, AP20187-mediated expansion of cells in dogs.
Nor, J.E., Hu, Y., Song, W., Spencer, D.M., and Nunez, G. (2002) Ablation of microvessels in vivo upon dimerization of iCaspase-9. Gene Ther 9:444-51.
AP20187-mediated oligomerization of caspase-9 triggers apoptosis of primary endothelial cells in vitro and in vivo.
Srinivasula, S.M., Poyet, J.L., Razmara, M., Datta, P., Zhang, Z., and Alnemri, E.S. (2002) The PYRIN-CARD Protein ASC Is an Activating Adaptor for Caspase-1. J Biol Chem 277:21119-22.
AP20187-mediated oligomerization of the caspase recruitment domain of ASC is sufficient to promote caspase-1 activation and secretion of IL-1beta
Welm, B.E., Freeman, K.W., Chen, M., Contreras, A., Spencer, D.M., and Rosen, J.M. (2002) Inducible dimerization of FGFR1: development of a mouse model to analyze progressive transformation of the mammary gland. J Cell Biol 157:703-14.
Uses AP20187-mediated oligomerization of FGFR1 to develop an inducible mouse model of breast cancer.
Xu, X., Tsvetkov, L.M., and Stern, D.F. (2002) Chk2 activation and phosphorylation-dependent oligomerization. Mol Cell Biol 22:4419-32.
AP20187-mediated oligomerization of Chk2, concomitant with limited DNA damage, is shown to increase Chk2 kinase activity.
Zhao, S., Zoller, K., Masuko, M., Rojnuckarin, P., Yang, X.O., Parganas, E., Kaushansky, K., Ihle, J.N., Papayannopoulou, T., Willerford, D.M., Clackson, T., and Blau, C.A. (2002) JAK2, complemented by a second signal from c-kit or flt-3, triggers extensive self-renewal of primary multipotential hemopoietic cells. Embo J 21:2159-67.
Demonstrates that AP20187-induced homodimerization and activation of a cytoplasmic JAK2 signaling domain, together with addition of either c-kit or flt-3 ligands, can trigger self-renewal of primary multipotential hemopoietic progenitor cells.

2001

Feng, H., Zeng, Y., Whitesell, L., and Katsanis, E. (2001) Stressed apoptotic tumor cells express heat shock proteins and elicit tumor-specific immunity. Blood 97:3505-12.
Uses AP20187-mediated dimerization of Fas to show that apoptotic tumor cells can be made immunogenic if they are heat-stressed prior to apoptotic induction
Iuliucci, J.D., Oliver, S.D., Morley, S., Ward, C., Ward, J., Dalgarno, D., Clackson, T., and Berger, H.J. (2001) Intravenous safety and pharmacokinetics of a novel dimerizer drug, AP1903, in healthy volunteers. J Clin Pharmacol 41:870-9.
AP1903 is shown to be safe and well tolerated in healthy human volunteers in a Phase 1 clinical trial.
Lucas, P.C., Yonezumi, M., Inohara, N., McAllister-Lucas, L.M., Abazeed, M.E., Chen, F.F., Yamaoka, S., Seto, M., and Nunez, G. (2001) Bcl10 and MALT1, independent targets of chromosomal translocation in malt lymphoma, cooperate in a novel NF-kappa B signaling pathway. J Biol Chem 276:19012-9.
AP1510-mediated oligomerization of the MALT1 caspase-like domain is sufficient to activate NF-kappaB.
McAllister-Lucas, L.M., Inohara, N., Lucas, P.C., Ruland, J., Benito, A., Li, Q., Chen, S., Chen, F.F., Yamaoka, S., Verma, I.M., Mak, T.W., and Nunez, G. (2001) Bimp1, a MAGUK family member linking protein kinase C activation to Bcl10-mediated NF-kappaB induction. J Biol Chem 276:30589-97.
A dominant negative Bimp1 is unable to block the ability of an AP1510-regulated MALT1 fusion protein to activate NF-kappaB, demonstrating the Bimp1 lies upstream on the signaling pathway.
Moulder, S.L., Yakes, F.M., Muthuswamy, S.K., Bianco, R., Simpson, J.F., and Arteaga, C.L. (2001) Epidermal growth factor receptor (HER1) tyrosine kinase inhibitor ZD1839 (Iressa) inhibits HER2/neu (erbB2)-overexpressing breast cancer cells in vitro and in vivo. Cancer Res 61:8887-95.
Uses AP1510-mediated activation of EGFR or HER2 receptor tyrosine kinases to demonstrate the relative specificity of the tyrosine kinase inhibitor Iressa(TM) for EGFR.
Muthuswamy, S.K., Li, D., Lelievre, S., Bissell, M.J., and Brugge, J.S. (2001) ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini. Nat Cell Biol 3:785-792.
Uses AP1510-mediated homodimerization to show that the ErbB1 and ErbB2 receptors differ in their ability to induce early stages of mammary carcinogenesis in vitro.
Neff, T., and Blau, C.A. (2001) Pharmacologically regulated cell therapy. Blood 97:2535-40.
Reviews the status and clinical potential of using small molecules to regulate the proliferation, differentiation or apoptosis of cells. The review focuses on the use of regulated homodimerization technology.
Nihalani, D., Meyer, D., Pajni, S., and Holzman, L.B. (2001) Mixed lineage kinase-dependent JNK activation is governed by interactions of scaffold protein JIP with MAPK module components. Embo J 20:3447-58.
AP20187-mediated dimerization of dual leucine zipper-bearing kinase (DLK) is sufficient to induce DLK autophosphorylation and JNK activation.
Ogura, Y., Inohara, N., Benito, A., Chen, F.F., Yamaoka, S., and Nunez, G. (2001) Nod2, a Nod1/Apaf-1 family member that is restricted to monocytes and activates NF-kappaB. J Biol Chem 276:4812-8.
AP1510-mediated oligomerization of Nod2 induces NF-kappaB activation.
Otto, K.G., Broudy, V.C., Lin, N.L., Parganas, E., Luthi, J.N., Drachman, J.G., Ihle, J.N., and Blau, C.A. (2001) Membrane localization is not required for Mpl function in normal hematopoietic cells. Blood 98:2077-83.
AP20187-mediated dimerization of mpl, targeted to the cytoplasm or plasma membrane, induces proliferation and differentiation of murine marrow cells in vitro.
Poyet, J.L., Srinivasula, S.M., and Alnemri, E.S. (2001) vCLAP, a caspase-recruitment domain-containing protein of equine Herpesvirus-2, persistently activates the Ikappa B kinases through oligomerization of IKKgamma. J Biol Chem 276:3183-7.
Uses AP1510 to demonstrate that vCLAP-induced oligomerization of IKKgamma is the triggering event leading to activation of the IKK complex.
Schotte, P., Van Loo, G., Carpentier, I., Vandenabeele, P., and Beyaert, R. (2001) Lithium sensitizes tumor cells in an NF-kB-independent way to caspase activation and apoptosis induced by tumor necrosis factor (TNF). Evidence for a role of the TNF receptor-associated death domain (TRADD) protein. J Biol Chem 276:25939-45.
AP1510-mediated oligomerization of TRADD signals cell death in a TNF-independent manner.
Shariat, S.F., Desai, S., Song, W., Khan, T., Zhao, J., Nguyen, C., Foster, B.A., Greenberg, N., Spencer, D.M., and Slawin, K.M. (2001) Adenovirus-mediated transfer of inducible caspases: a novel "death switch" gene therapeutic approach to prostate cancer. Cancer Res 61:2562-71.
Demonstrates that AP20187-mediated oligomerization of caspase-1 delivered by adenovirus vectors induces apoptosis and reduces tumor growth in a murine prostate cancer model.
Smith, K.M., and Van Etten, R.A. (2001) Activation of c-Abl kinase activity and transformation by a chemical inducer of dimerization. J Biol Chem 276:24372-9.
AP1510-mediated dimerization is sufficient to activate Abl+ kinase activity and transformation.
Soldevila, G., Castellanos, C., Malissen, M., and Berg, L.J. (2001) Analysis of the Individual Role of the TCRzeta Chain in Transgenic Mice after Conditional Activation with Chemical Inducers of Dimerization. Cell Immunol 214:123-38.
AP1510-mediated oligomerization of the cytoplasmic tail of the T cell receptor zeta chain is sufficient to induce tyrosine phosphorylation but not a full functional response in primary T lymphocytes or thymocytes.
Thomis, D.C., Marktel, S., Bonini, C., Traversari, C., Gilman, M., Bordignon, C., and Clackson, T. (2001) A Fas-based suicide switch in human T cells for the treatment of graft-versus-host disease. Blood 97:1249-57.
Describes optimization and preclinical testing of ARIAD's inducible Fas system for use as a cellular "suicide switch" for the treatment of graft-vs-host disease. A Fas construct linked to a cell surface marker is introduced into primary human T cells using a retroviral vector. Low doses of AP1903 are shown to rapidly and efficiently eliminate the engineered cells, independently of their activation status.
Wang, J., Chun, H.J., Wong, W., Spencer, D.M., and Lenardo, M.J. (2001) Caspase-10 is an initiator caspase in death receptor signaling. Proc Natl Acad Sci USA 98:13884-8.
AP20187-mediated dimerization of caspase-10 induces apoptosis.
Whitney, M.L., Otto, K.G., Blau, C.A., Reinecke, H., and Murry, C.E. (2001) Control of myoblast proliferation with a synthetic ligand. J Biol Chem 276:41191-6.
AP20187-mediated dimerization of the FGFR-1 cytoplasmic domain reversibly stimulates myoblast proliferation and blocks differentiation into myotubes.
Xie, X., Zhao, X., Liu, Y., Zhang, J., Matusik, R.J., Slawin, K.M., and Spencer, D.M. (2001) Adenovirus-mediated tissue-targeted expression of a caspase-9-based artificial death switch for the treatment of prostate cancer. Cancer Res 61:6795-804.
AP20187-mediated oligomerization of caspase-9 delivered to xenograft prostate tumors in mice using adenoviral vectors induces apoptosis and significantly increases host survival.
Xu, X., Tsvetkov, L.M., and Stern, D.F. (2001) Chk2 activation and phosphorylation-dependent oligomerization. Mol Cell Biol 22:4419-32.
Demonstrates that AP20187-mediated oligomerization of CHK2, concomitant with limited DNA damage augments Chk2 kinase activity.
Zeng, H., Masuko, M., Jin, L., Neff, T., Otto, K.G., and Blau, C.A. (2001) Receptor specificity in the self-renewal and differentiation of primary multipotential hemopoietic cells. Blood 98:328-34.
FK1012 or AP20187-mediated oligomerization of Mpl, the G-CSF receptor or Flt-3 is sufficient to induce growth of BaF3 cells, but only dimerization of Mpl induces mouse hematopoietic cell self renewal.
Zhao, T., and Newman, P.J. (2001) Integrin Activation by Regulated Dimerization and Oligomerization of Platelet Endothelial Cell Adhesion Molecule (PECAM)-1 from Within the Cell. J Cell Biol 152:65-74.
Uses AP1510-mediated dimerization of PECAM-1 to study the influence of its oligomerization state on integrin activation.

2000

Bayly, R., and LeBrun, D.P. (2000) Role for homodimerization in growth deregulation by E2a fusion proteins. Mol Cell Biol 20:5789-96.
AP20187-mediated dimerization of a portion of E2a is sufficient to promote growth deregulation.
Chadli, A., Bouhouche, I., Sullivan, W., Stensgard, B., McMahon, N., Catelli, M.G., and Toft, D.O. (2000) Dimerization and N-terminal domain proximity underlie the function of the molecular chaperone heat shock protein 90. Proc Natl Acad Sci USA 97:12524-9.
Uses AP20187 to demonstrate that dimerization of hsp90 is required for p23 binding.
Inohara, N., Koseki, T., Lin, J., del Peso, L., Lucas, P.C., Chen, F.F., Ogura, Y., and Nunez, G. (2000) An induced proximity model for NF-kappaB activation in the Nod1/RICK and RIP signaling pathways. J Biol Chem 275:27823-31.
AP1510-mediated oligomerization of Nod1, RICK, RIP and IKK-alpha, -beta, or gamma is sufficient to activate NF-kappaB.
Jin, L., Zeng, H., Chien, S., Otto, K.G., Richard, R.E., Emery, D.W., and Blau, C.A. (2000) In vivo selection using a cell-growth switch. Nat Genet 26:64-6.
The first in vivo demonstration of a generalized platform for conditionally expanding an engineered cell population. AP20187-mediated dimerization of the signaling domain of mpl is shown to be sufficient to induce expansion and differentiation of engineered bone marrow cells in a mouse.
Kopytek, S.J., Standaert, R.F., Dyer, J.C., and Hu, J.C. (2000) Chemically induced dimerization of dihydrofolate reductase by a homobifunctional dimer of methotrexate. Chem Biol 7:313-321.
Describes the development of the alternative dimerizer bisMTX, a dimeric version of the DHFR ligand methotrexate, and demonstrates its ability to dimerize DHFR in vitro.
Lesley, J., English, N., Charles, C., and Hyman, R. (2000) The role of the CD44 cytoplasmic and transmembrane domains in constitutive and inducible hyaluronan binding. Eur J Immunol 30:245-53.
Uses AP1510 to explore the role of dimerization in the hyaluronan binding activity of CD44.
Matsumura, H., Shimizu, Y., Ohsawa, Y., Kawahara, A., Uchiyama, Y., and Nagata, S. (2000) Necrotic death pathway in Fas receptor signaling. J Cell Biol 151:1247-56.
Uses AP1510-mediated oligomerization of the death effector domain of FADD to demonstrate its role in the FADD-mediated necrotic death pathway.
Poyet, J.L., Srinivasula, S.M., Lin, J.H., Fernandes-Alnemri, T., Yamaoka, S., Tsichlis, P.N., and Alnemri, E.S. (2000) Activation of the Ikappa B kinases by RIP via IKKgamma /NEMO-mediated oligomerization. J Biol Chem 275:37966-77.
Uses AP1510 to demonstrate the role of oligomerization in the activation of IKK.
Richard, R.E., Wood, B., Zeng, H., Jin, L., Papayannopoulou, T., and Blau, C.A. (2000) Expansion of genetically modified primary human hemopoietic cells using chemical inducers of dimerization. Blood 95:430-6.
AP1903-mediated dimerization of the signaling domain of mpl stimulates proliferation of engineered primary human hematopoietic cells in vitro.
Schlake, T., Schorpp, M., Maul-Pavicic, A., Malashenko, A.M., and Boehm, T. (2000) Forkhead/winged-helix transcription factor whn regulates hair keratin gene expression: molecular analysis of the nude skin phenotype. Dev Dyn 217:368-76.
AP1510 mediated dimerization of DNA binding and transcriptional activation domains from the Whn transcription factor activates transcription of a target gene.
Spencer, D.M. (2000) Developments in suicide genes for preclinical and clinical applications. Curr Opin Mol Ther 2:433-40.
Reviews the potential clinical application of suicide genes with an emphasis on those regulated by dimerizers (e.g. Fas and caspases).
Wang, J.J., Rabizadeh, S., Tasinato, A., Sperandio, S., Ye, X., Green, M., Assa-Munt, N., Spencer, D., and Bredesen, D.E. (2000) Dimerization-dependent block of the proapoptotic effect of p75(NTR). J Neurosci Res 60:587-93.
Uses AP1510 to demonstrate the role of dimerization in inhibiting the proapoptotic effect of the common neurotrophin receptor p75(NTR).
Yang, W., Rozamus, L.W., Narula, S., Rollins, C.T., Yuan, R., Andrade, L.J., Ram, M.K., Phillips, T.B., van Schravendijk, M.R., Dalgarno, D., Clackson, T., and Holt, D.A. (2000) Investigating protein-ligand interactions with a mutant FKBP possessing a designed specificity pocket. J Med Chem 43:1135-42.
Describes a large number of synthetic AP1903-like ligands designed to bind to the F36V mutant FKBP.

1999

Amara, J.F., Courage, N.L., and Gilman, M. (1999) Cell surface tagging and a suicide mechanism in a single chimeric human protein. Hum Gene Ther 10:2651-5.
Describes the development of a humanized, Fas-based suicide switch in which the Fas fusion protein is covalently linked to an extracellular cell surface marker, LNGFR. AP1903-mediated oligomerization of Fas induces apoptosis.
Baud, V., Liu, Z.G., Bennett, B., Suzuki, N., Xia, Y., and Karin, M. (1999) Signaling by proinflammatory cytokines: oligomerization of TRAF2 and TRAF6 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain. Genes Dev 13:1297-308.
FK1012-mediated oligomerization of TRAF2 or TRAF6 is sufficient to activate multiple downstream targets including JNK and IKK.
Blau, C.A. (1999) In vivo selection of genetically modified bone marrow cells. Prog Exp Tumor Res 36:162-71.
Reviews the potential for dimerizer-mediated proliferation of engineered cells to overcome the limitations associated with inefficient gene transfer into bone marrow cells.
Cheng, H.H., Xu, L., Kumagai, H., and Simoni, R.D. (1999) Oligomerization state influences the degradation rate of 3-hydroxy-3- methylglutaryl-CoA reductase. J Biol Chem 274:17171-8.
Uses AP1510-mediated dimerization of HMGR to study the influence of its oligomerization state on the rate of degradation.
Fan, L., Freeman, K.W., Khan, T., Pham, E., and Spencer, D.M. (1999) Improved artificial death switches based on caspases and FADD. Hum Gene Ther 10:2273-85.
AP1903-mediated activation of the signaling domains of Fas, FADD or caspase 1, 3, 8 or 9 induces apoptosis. Potency is optimized based on the extent of oligomerization, the spacing between FKBP and signaling domains and intracellular localization.
Lee, S.F., Huang, H.M., Chao, J.R., Lin, S., Yang-Yen, H.F., and Yen, J.J. (1999) Cytokine receptor common beta chain as a potential activator of cytokine withdrawal-induced apoptosis. Mol Cell Biol 19:7399-409.
Uses AP1510 to explore the effect of dimerization on the death promoting activity of the cytokine receptor common beta chain.
Muthuswamy, S.K., Gilman, M., and Brugge, J.S. (1999) Controlled dimerization of ErbB receptors provides evidence for differential signaling by homo- and heterodimers. Mol Cell Biol 19:6845-57.
Uses the homodimerizer AP1510 and the heterodimerizer rapamycin to explore the differential signaling activities of ErbB1 and ErbB2 homodimers and heterodimers.

1998

Clackson, T. (1998) Redesigning small molecule-protein interfaces. Curr Opin Struct Biol 8:451-8.
Reviews examples of redesigned small molecule-protein interfaces, with an emphasis on the remodeling of the interface between FKBP and synthetic ligands (the basis of the homodimerizers AP1903 and AP20187).
Clackson, T., Yang, W., Rozamus, L.W., Hatada, M., Amara, J.F., Rollins, C.T., Stevenson, L.F., Magari, S.R., Wood, S.A., Courage, N.L., Lu, X., Cerasoli, F., Jr., Gilman, M., and Holt, D.A. (1998) Redesigning an FKBP-ligand interface to generate chemical dimerizers with novel specificity. Proc Natl Acad Sci USA 95:10437-42.
Describes the development of a third generation FKBP homodimerizer, AP1903, which binds specifically to a mutated FKBP over the wild type protein. AP1903-mediated oligomerization of a Fas fusion protein is shown to induce apoptosis in engineered cells in vitro, and in vivo in implanted mice.
De Angelis, D.A., Miesenbock, G., Zemelman, B.V., and Rothman, J.E. (1998) PRIM: proximity imaging of green fluorescent protein-tagged polypeptides. Proc Natl Acad Sci USA 95:12312-6.
Uses FK1012-mediated homodimerization of GFP to demonstrate the ability of PRIM to detect protein-protein interactions in cells.
Gross, A., Jockel, J., Wei, M.C., and Korsmeyer, S.J. (1998) Enforced dimerization of BAX results in its translocation, mitochondrial dysfunction and apoptosis. Embo J 17:3878-85.
FK1012-mediated dimerization of BAX is sufficient to induce apoptosis.
Hato, T., Pampori, N., and Shattil, S.J. (1998) Complementary roles for receptor clustering and conformational change in the adhesive and signaling functions of integrin alphaIIb beta3. J Cell Biol 141:1685-95.
Uses AP1510 to study the effects of clustering the integrin alphaIIbbeta3.
Hu, Y., Ding, L., Spencer, D.M., and Nunez, G. (1998) WD-40 repeat region regulates Apaf-1 self-association and procaspase-9 activation. J Biol Chem 273:33489-94.
Uses FK1012-mediated oligomerization to explore the mechanism by which Apaf-1 activates procaspase-9.
Jin, L., Asano, H., and Blau, C.A. (1998) Stimulating cell proliferation through the pharmacologic activation of c-kit. Blood 91:890-7.
AP1510-mediated dimerization of the signaling domain of the c-kit receptor is sufficient to stimulate proliferation of engineered cells in vitro.
Jin, L., Siritanaratkul, N., Emery, D.W., Richard, R.E., Kaushansky, K., Papayannopoulou, T., and Blau, C.A. (1998) Targeted expansion of genetically modified bone marrow cells. Proc Natl Acad Sci USA 95:8093-7.
FK1012-mediated dimerization of the signaling domain of mpl stimulates the reversible expansion of engineered bone marrow cells in vitro.
Kawahara, A., Ohsawa, Y., Matsumura, H., Uchiyama, Y., and Nagata, S. (1998) Caspase-independent cell killing by Fas-associated protein with death domain. J Cell Biol 143:1353-60.
FK1012-mediated oligomerization of FADD reveals two separate pathways by which FADD induces apoptosis.
Keenan, T., Yaeger, D.R., Courage, N.L., Rollins, C.T., Pavone, M.E., Rivera, V.M., Yang, W., Guo, T., Amara, J.F., Clackson, T., Gilman, M., and Holt, D.A. (1998) Synthesis and activity of bivalent FKBP12 ligands for the regulated dimerization of proteins. Bioorg Med Chem 6:1309-35.
Describes the synthesis and activities of an extended set of derivatives of the dimerizer AP1510.
MacCorkle, R.A., Freeman, K.W., and Spencer, D.M. (1998) Synthetic activation of caspases: artificial death switches. Proc Natl Acad Sci USA 95:3655-60.
Uses AP1903 to demonstrate that oligomerization of caspase-1 or -3 is sufficient to trigger apoptosis in a Bcl-xL -independent manner.
Muzio, M., Stockwell, B.R., Stennicke, H.R., Salvesen, G.S., and Dixit, V.M. (1998) An induced proximity model for caspase-8 activation. J Biol Chem 273:2926-30.
FK1012-mediated oligomerization of caspase-8 is sufficient to induce apoptosis.
Schultz, L.W., and Clardy, J. (1998) Chemical inducers of dimerization: the atomic structure of FKBP12- FK1012A-FKBP12. Bioorg Med Chem Lett 8:1-6.
Describes the high resolution x-ray structure of FK1012 bound to two molecules of FKBP.
Stockwell, B.R., and Schreiber, S.L. (1998) Probing the role of homomeric and heteromeric receptor interactions in TGF-beta signaling using small molecule dimerizers. Curr Biol 8:761-70.
Explores the role of receptor oligomerization in TGF-beta signaling using a variety of homo- and heterodimerizers including rapamycin and FK1012.
Yang, J., Symes, K., Mercola, M., and Schreiber, S.L. (1998) Small-molecule control of insulin and PDGF receptor signaling and the role of membrane attachment. Curr Biol 8:11-8.
FK1012-mediated oligomerization of the PDGF and insulin receptors is sufficient to induce signaling. Activation of the PDGF receptor in Xenopus embryos induced the formation of mesoderm.
Yang, X., Chang, H.Y., and Baltimore, D. (1998) Essential role of CED-4 oligomerization in CED-3 activation and apoptosis. Science 281:1355-7.
AP1510-mediated oligomerization of CED-3 is sufficient to induce apoptosis.
Yang, X., Chang, H.Y., and Baltimore, D. (1998) Autoproteolytic activation of pro-caspases by oligomerization. Mol Cell 1:319-25.
AP1510-mediated oligomerization of pro-caspase-8 is sufficient to induce apoptosis.

1997

Amara, J.F., Clackson, T., Rivera, V.M., Guo, T., Keenan, T., Natesan, S., Pollock, R., Yang, W., Courage, N.L., Holt, D.A., and Gilman, M. (1997) A versatile synthetic dimerizer for the regulation of protein-protein interactions. Proc Natl Acad Sci USA 94:10618-23.
Describes the development of the second generation, wholly synthetic FKBP dimerizer, AP1510. The synthetic nature of AP1510 makes it more amenable to modification and more readily synthesized in larger quantities. AP1510-mediated oligomerization of a Fas receptor and an engineered transcription factor are shown to induce apoptosis and transcription of a target gene, respectively.
Blau, C.A., Peterson, K.R., Drachman, J.G., and Spencer, D.M. (1997) A proliferation switch for genetically modified cells. Proc Natl Acad Sci USA 94:3076-81.
The first demonstration of the use of dimerizers to specifically and reversibly stimulate the proliferation of a population of engineered cells. FK1012-mediated dimerization of the signaling domain of the erythropoietin receptor is shown to stimulate the proliferation of cells normally dependent on IL-3 for growth.
Freiberg, R.A., Spencer, D.M., Choate, K.A., Duh, H.J., Schreiber, S.L., Crabtree, G.R., and Khavari, P.A. (1997) Fas signal transduction triggers either proliferation or apoptosis in human fibroblasts. J Invest Dermatol 108:215-9.
Examines the effects of FK1012-mediated oligomerization of Fas in fibroblasts.
Gires, O., Zimber-Strobl, U., Gonnella, R., Ueffing, M., Marschall, G., Zeidler, R., Pich, D., and Hammerschmidt, W. (1997) Latent membrane protein 1 of Epstein-Barr virus mimics a constitutively active receptor molecule. Embo J 16:6131-40.
Uses AP1510-mediated oligomerization to explore the natural function of LMP1.
Graef, I.A., Holsinger, L.J., Diver, S., Schreiber, S.L., and Crabtree, G.R. (1997) Proximity and orientation underlie signaling by the non-receptor tyrosine kinase ZAP70. Embo J 16:5618-28.
Explores the effects of membrane recruitment, orientation and oligomerization of ZAP-70 using the homodimerizer FK1012 and the heterodimerizer rapamycin.
Kieser, A., Kilger, E., Gires, O., Ueffing, M., Kolch, W., and Hammerschmidt, W. (1997) Epstein-Barr virus latent membrane protein-1 triggers AP-1 activity via the c-Jun N-terminal kinase cascade. Embo J 16:6478-85.
AP1510-mediated oligomerization of LMP-1 induces JNK1 activity.
Yap, A.S., Brieher, W.M., Pruschy, M., and Gumbiner, B.M. (1997) Lateral clustering of the adhesive ectodomain: a fundamental determinant of cadherin function. Curr Biol 7:308-15.
FK1012-induced oligomerization of C-cadherin increases cellular adhesion.

1996

Belshaw, P.J., Spencer, D.M., Crabtree, G.R., and Schreiber, S.L. (1996) Controlling programmed cell death with a cyclophilin-cyclosporin-based chemical inducer of dimerization. Chem Biol 3:731-8.
Oligomerization of Fas with a cyclophilin-cyclosporin-based homodimerizer system induces apoptosis.
Crabtree, G.R., and Schreiber, S.L. (1996) Three-part inventions: intracellular signaling and induced proximity. Trends Biochem Sci 21:418-22.
Reviews the potential of dimerizers to control biological processes.
Farrar, M.A., Alberol, I., and Perlmutter, R.M. (1996) Activation of the Raf-1 kinase cascade by coumermycin-induced dimerization. Nature 383:178-81.
Describes the development of the alternative dimerizer coumermycin, a gyrase B ligand, and demonstrates that oligomerization of Raf is sufficient to activate downstream signaling. The coumermycin dimerizer system has been used in several other studies to induce activation of a variety of signaling proteins. References describing these studies are listed at the end of this section.
Freiberg, R.A., Spencer, D.M., Choate, K.A., Peng, P.D., Schreiber, S.L., Crabtree, G.R., and Khavari, P.A. (1996) Specific triggering of the Fas signal transduction pathway in normal human keratinocytes. J Biol Chem 271:31666-9.
FK1012-mediated oligomerization of Fas is sufficient to trigger apoptosis in keratinocytes.
Luo, Z., Tzivion, G., Belshaw, P.J., Vavvas, D., Marshall, M., and Avruch, J. (1996) Oligomerization activates c-Raf-1 through a Ras-dependent mechanism. Nature 383:181-5.
FK1012-mediated oligomerization of Raf is sufficient to induce its activity through a Ras-dependent mechanism.
Spencer, D.M. (1996) Creating conditional mutations in mammals. Trends Genet 12:181-7.
Reviews the various approaches that can be used to conditionally express or control the activity of proteins in mammals, with an emphasis on the use of dimerizers.
Spencer, D.M., Belshaw, P.J., Chen, L., Ho, S.N., Randazzo, F., Crabtree, G.R., and Schreiber, S.L. (1996) Functional analysis of Fas signaling in vivo using synthetic inducers of dimerization. Curr Biol 6:839-47.
FK1012-mediated oligomerization of Fas is sufficient to induce apoptosis in vitro, and in trangenic mice engineered to express the chimeric Fas in thymocytes.

1995

Holsinger, L.J., Spencer, D.M., Austin, D.J., Schreiber, S.L., and Crabtree, G.R. (1995) Signal transduction in T lymphocytes using a conditional allele of Sos. Proc Natl Acad Sci USA 92:9810-4.
FK1012-mediated recruitment of Sos to the plasma membrane is sufficient to activate Ras in T cells.
Spencer, D.M., Graef, I., Austin, D.J., Schreiber, S.L., and Crabtree, G.R. (1995) A general strategy for producing conditional alleles of Src-like tyrosine kinases. Proc Natl Acad Sci USA 92:9805-9.
FK1012-mediated recruitment of cytoplasmic Src to the plasma membrane is sufficient to induce signaling.

1994

Austin, D.J., Crabtree, G.R., and Schreiber, S.L. (1994) Proximity versus allostery: the role of regulated protein dimerization in biology. Chem Biol 1:131-6.
Reviews the natural role of protein dimerization in cellular processes and describes how dimerizers provide a general method to regulate such processes.
Pruschy, M.N., Spencer, D.M., Kapoor, T.M., Miyake, H., Crabtree, G.R., and Schreiber, S.L. (1994) Mechanistic studies of a signaling pathway activated by the organic dimerizer FK1012. Chem Biol 1:163-72.
Oligomerization of the T-cell receptor-zeta chain with FK1012 is sufficient to induce downstream signaling.

1993

Spencer, D.M., Wandless, T.J., Schreiber, S.L., and Crabtree, G.R. (1993) Controlling signal transduction with synthetic ligands. Science 262:1019-24.
The first description of the use of dimerizers. Describes the development of FK1012, a semisynthetic dimer of FK506 that can simultaneously bind two molecules of FKBP12. FK1012-mediated oligomerization of the T cell receptor-zeta chain is shown to be sufficient to induce signaling.

ARGENT Regulated Homodimerization Using Coumermycin

An alternative dimerization system has been described that uses the natural product coumermycin and its binding proteins. Although ARIAD does not distribute reagents for this system, publications that describe its use are listed below.