Vance, Keith W., Shaw, Heather M., Rodriguez, Mercedes, Ott, Sascha and Goding, Colin R.. (2010) The retinoblastoma protein modulates Tbx2 functional specificity. Molecular Biology of the Cell, Vol.21 (No.15). pp. 2770-2779. ISSN 1059-1524

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Abstract

Tbx2 is a member of a large family of transcription factors defined by homology to the T-box DNA-binding domain. Tbx2 plays a key role in embryonic development, and in cancer through its capacity to suppress senescence and promote invasiveness. Despite its importance, little is known of how Tbx2 is regulated or how it achieves target gene specificity. Here we show that Tbx2 specifically associates with active hypophosphorylated retinoblastoma protein (Rb1), a known regulator of many transcription factors involved in cell cycle progression and cellular differentiation, but not with the Rb1-related proteins p107 or p130. The interaction with Rb1 maps to a domain immediately carboxy-terminal to the T-box and enhances Tbx2 DNA binding and transcriptional repression. Microarray analysis of melanoma cells expressing inducible dominant-negative Tbx2, comprising the T-box and either an intact or mutated Rb1 interaction domain, shows that Tbx2 regulates the expression of many genes involved in cell cycle control and that a mutation which disrupts the Rb1-Tbx2 interaction also affects Tbx2 target gene selectivity. Taken together, the data show that Rb1 is an important determinant of Tbx2 functional specificity.

Item Type:	Journal Article
Subjects:	Q Science > QP Physiology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010) Faculty of Science > Centre for Systems Biology
Library of Congress Subject Headings (LCSH):	Transcription factors, Retinoblastoma, Gene expression
Journal or Publication Title:	Molecular Biology of the Cell
Publisher:	American Society for Cell Biology
ISSN:	1059-1524
Date:	1 August 2010
Volume:	Vol.21
Number:	No.15
Number of Pages:	10
Page Range:	pp. 2770-2779
Identification Number:	10.1091/mbc.E09-12-1029
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access
Funder:	Medical Research Council (Great Britain) (MRC), University of Warwick, Wellcome Trust (London, England)
References:	Abrahams, A., Mowla, S., Parker, M. I., Goding, C. R., and Prince, S. (2008). UV-mediated regulation of the anti-senescence factor Tbx2. J. Biol. Chem. 283, 2223–2230. Aksan, I., and Goding, C. R. (1998). Targeting the microphthalmia basic helix-loop-helix-leucine zipper transcription factor to a subset of E-box elements in vitro and in vivo. Mol. Cell. Biol. 18, 6930–6938. Amellem, O., Stokke, T., Sandvik, J. A., and Pettersen, E. O. (1996). The retinoblastoma gene product is reversibly dephosphorylated and bound in the nucleus in S and G2 phases during hypoxic stress. Exp. Cell Res. 227, 106–115. Bandyopadhyay, D., and Medrano, E. E. (2000). Melanin accumulation accelerates melanocyte senescence by a mechanism involving p16INK4a/CDK4/ pRB and E2F1. Ann. NY Acad. Sci. 908, 71–84. Bilican, B., and Goding, C. R. (2006). Cell cycle regulation of the T-box transcription factor tbx2. Exp. Cell Res. 312, 2358–2366. Boogerd, K. J., Wong, L. Y., Christoffels, V. M., Klarenbeek, M., Ruijter, J. M., Moorman, A. F., and Barnett, P. (2008). Msx1 and Msx2 are functional interacting partners of T-box factors in the regulation of connexin43. Cardiovasc. Res. 78, 485–493. Brehm, A., and Kouzarides, T. (1999). Retinoblastoma protein meets chromatin. Trends Biochem. Sci. 24, 142–145. Brehm, A., Miska, E. A., McCance, D. J., Reid, J. L., Bannister, A. J., and Kouzarides, T. (1998). Retinoblastoma protein recruits histone deacetylase to repress transcription. Nature 391, 597–601. Butz, N. V., Campbell, C. E., and Gronostajski, R. M. (2004). Differential target gene activation by TBX2 and TBX2VP16, evidence for activation domaindependent modulation of gene target specificity. Gene 342, 67–76. Carlson, H., Ota, S., Song, Y., Chen, Y., and Hurlin, P. J. (2002). Tbx3 impinges on the p53 pathway to suppress apoptosis, facilitate cell transformation and block myogenic differentiation. Oncogene 21, 3827–3835. Carreira, S., Dexter, T. J., Yavuzer, U., Easty, D. J., and Goding, C. R. (1998). Brachyury-related transcription factor Tbx2 and repression of the melanocytespecific TRP-1 promoter. Mol. Cell. Biol. 18, 5099–5108. Carreira, S., Goodall, J., Aksan, I., La Rocca, S. A., Galibert, M. D., Denat, L., Larue, L., and Goding, C. R. (2005). Mitf cooperates with Rb1 and activates p21Cip1 expression to regulate cell cycle progression. Nature 433, 764–769. Chen, J., Zhong, Q., Wang, J., Cameron, R. S., Borke, J. L., Isales, C. M., and Bollag, R. J. (2001). Microarray analysis of Tbx2-directed gene expression: a possible role in osteogenesis. Mol. Cell. Endocrinol. 177, 43–54. Chen, J. R., Chatterjee, B., Meyer, R., Yu, J. C., Borke, J. L., Isales, C. M., Kirby, M. L., Lo, C. W., and Bollag, R. J. (2004). Tbx2 represses expression of Connexin43 in osteoblastic-like cells. Calcif. Tissue Int. 74, 561–573. Chen, P. L., Riley, D. J., Chen-Kiang, S., and Lee, W. H. (1996a). Retinoblastoma protein directly interacts with and activates the transcription factor NF-IL6. Proc. Natl. Acad. Sci. USA 93, 465–469. Chen, P. L., Riley, D. J., Chen, Y., and Lee, W. H. (1996b). Retinoblastoma protein positively regulates terminal adipocyte differentiation through direct interaction with C/EBPs. Genes Dev. 10, 2794–2804. Conlon, F. L., Fairclough, L., Price, B. M., Casey, E. S., and Smith, J. C. (2001). Determinants of T box protein specificity. Development 128, 3749–3758. Dai, M., et al. (2005). Evolving gene/transcript definitions significantly alter the interpretation of GeneChip data. Nucleic Acids Res. 33, e175. Davis, E., Teng, H., Bilican, B., Parker, M. I., Liu, B., Carriera, S., Goding, C. R., and Prince, S. (2007). Ectopic Tbx2 expression results in polyploidy and cisplatin resistance. Oncogene 27, 976–984. Demay, F., Bilican, B., Rodriguez, M., Carreira, S., Pontecorvi, M., Ling, Y., and Goding, C. R. (2007). T-box factors: targeting to chromatin and interaction with the histone H3 N-terminal tail. Pigment Cell Res. 20, 279–287. Dunaief, J. L., Strober, B. E., Guha, S., Khavari, P. A., Alin, K., Luban, J., Begemann, M., Crabtree, G. R., and Goff, S. P. (1994). The retinoblastoma protein and BRG1 form a complex and cooperate to induce cell cycle arrest. Cell 79, 119–130. Fajas, L., Paul, C., Zugasti, O., Le Cam, L., Polanowska, J., Fabbrizio, E., Medema, R., Vignais, M. L., and Sardet, C. (2000). pRB binds to and modulates the transrepressing activity of the E1A-regulated transcription factor p120E4F. Proc. Natl. Acad. Sci. USA 97, 7738–7743. Gentleman, R. C., et al. (2004). Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80. Harrelson, Z., Kelly, R. G., Goldin, S. N., Gibson-Brown, J. J., Bollag, R. J., Silver, L. M., and Papaioannou, V. E. (2004). Tbx2 is essential for patterning the atrioventricular canal and for morphogenesis of the outflow tract during heart development. Development 131, 5041–5052. Herrmann, B. G., Labeit, S., Poustka, A., King, T. R., and Lehrach, H. (1990). Cloning of the T gene required in mesoderm formation in the mouse. Nature 343, 617–622. Hiroi, Y., Kudoh, S., Monzen, K., Ikeda, Y., Yazaki, Y., Nagai, R., and Komuro, I. (2001). Tbx5 associates with Nkx2-5 and synergistically promotes cardiomyocyte differentiation. Nat. Genet. 28, 276–280. Hitachi, K., Danno, H., Tazumi, S., Aihara, Y., Uchiyama, H., Okabayashi, K., Kondow, A., and Asashima, M. (2009). The Xenopus Bowline/Ripply family proteins negatively regulate the transcriptional activity of T-box transcription factors. Int. J. Dev. Biol. 53, 631–639. Hoek, K., et al. (2004). Expression profiling reveals novel pathways in the transformation of melanocytes to melanomas. Cancer Res. 64, 5270–5282. Ishida, S., Huang, E., Zuzan, H., Spang, R., Leone, G., West, M., and Nevins, J. R. (2001). Role for E2F in control of both DNA replication and mitotic functions as revealed from DNA microarray analysis. Mol. Cell. Biol. 21, 4684–4699. Jacobs, J. J., et al. (2000). Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19(ARF)) and is amplified in a subset of human breast cancers. Nat. Genet. 26, 291–299. Jonsson, G., Dahl, C., Staaf, J., Sandberg, T., Bendahl, P. O., Ringner, M., Guldberg, P., and Borg, A. (2007). Genomic profiling of malignant melanoma using tiling-resolution arrayCGH. Oncogene 26, 4738–4748. Kawamura, A., Koshida, S., and Takada, S. (2008). Activator-to-repressor conversion of T-box transcription factors by the Ripply family of Groucho/ TLE-associated mediators. Mol. Cell. Biol. 28, 3236–3244. Lamolet, B., Pulichino, A. M., Lamonerie, T., Gauthier, Y., Brue, T., Enjalbert, A., and Drouin, J. (2001). A pituitary cell-restricted T box factor, Tpit, activates POMC transcription in cooperation with Pitx homeoproteins. Cell 104, 849– 859. Lavender, P., Vandel, L., Bannister, A. J., and Kouzarides, T. (1997). The HMG-box transcription factor HBP1 is targeted by the pocket proteins and E1A. Oncogene 14, 2721–2728. Lewis, M. D., Miller, S. A., Miazgowicz, M. M., Beima, K. M., and Weinmann, A. S. (2007). T-bet’s ability to regulate individual target genes requires the conserved T-box domain to recruit histone methyltransferase activity and a separate family member-specific transactivation domain. Mol. Cell. Biol. 27, 8510–8521. Lingbeek, M. E., Jacobs, J. J., and van Lohuizen, M. (2002). The T-box repressors TBX2 and TBX3 specifically regulate the tumor suppressor gene p14ARF via a variant T-site in the initiator. J. Biol. Chem. 277, 26120–26127. Linke, S. P., Harris, M. P., Neugebauer, S. E., Clarkin, K. C., Shepard, H. M., Maneval, D. C., and Wahl, G. M. (1997). p53-mediated accumulation of hypophosphorylated pRb after the G1 restriction point fails to halt cell cycle progression. Oncogene 15, 337–345. Lipinski, M. M., and Jacks, T. (1999). The retinoblastoma gene family in differentiation and development. Oncogene 18, 7873–7882. Lomnytska, M., Dubrovska, A., Hellman, U., Volodko, N., and Souchelnytskyi, S. (2006). Increased expression of cSHMT, Tbx3 and utrophin in plasma of ovarian and breast cancer patients. Int. J. Cancer 118, 412–421. Lyng, H., Brovig, R. S., Svendsrud, D. H., Holm, R., Kaalhus, O., Knutstad, K., Oksefjell, H., Sundfor, K., Kristensen, G. B., and Stokke, T. (2006). Gene expressions and copy numbers associated with metastatic phenotypes of uterine cervical cancer. BMC Genom. 7, 268. Magnaghi-Jaulin, L., Groisman, R., Naguibneva, I., Robin, P., Lorain, S., Le Villain, J. P., Troalen, F., Trouche, D., and Harel-Bellan, A. (1998). Retinoblastoma protein represses transcription by recruiting a histone deacetylase. Nature 391, 601–605. Mahlamaki, E. H., Barlund, M., Tanner, M., Gorunova, L., Hoglund, M., Karhu, R., and Kallioniemi, A. (2002). Frequent amplification of 8q24, 11q, 17q, and 20q-specific genes in pancreatic cancer. Genes Chromosomes Cancer 35, 353–358. Maira, M., Couture, C., Le Martelot, G., Pulichino, A. M., Bilodeau, S., and Drouin, J. (2003). The T-box factor Tpit recruits SRC/p160 co-activators and mediates hormone action. J. Biol. Chem. 278, 46523–46532. Markey, M. P., Angus, S. P., Strobeck, M. W., Williams, S. L., Gunawardena, R. W., Aronow, B. J., and Knudsen, E. S. (2002). Unbiased analysis of RBmediated transcriptional repression identifies novel targets and distinctions from E2F action. Cancer Res. 62, 6587–6597. Martini, E., Roche, D. M., Marheineke, K., Verreault, A., and Almouzni, G. (1998). Recruitment of phosphorylated chromatin assembly factor 1 to chromatin after UV irradiation of human cells. J. Cell Biol. 143, 563–575. Melnikova, V. O., Bolshakov, S. V., Walker, C., and Ananthaswamy, H. N. (2004). Genomic alterations in spontaneous and carcinogen-induced murine melanoma cell lines. Oncogene 23, 2347–2356. Messenger, N. J., Kabitschke, C., Andrews, R., Grimmer, D., Nunez Miguel, R., Blundell, T. L., Smith, J. C., and Wardle, F. C. (2005). Functional specificity of the Xenopus T-domain protein Brachyury is conferred by its ability to interact with Smad1. Dev. Cell 8, 599–610. Michaloglou, C., Vredeveld, L. C., Soengas, M. S., Denoyelle, C., Kuilman, T., van der Horst, C. M., Majoor, D. M., Shay, J. W., Mooi, W. J., and Peeper, D. S. (2005). BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724. Miller, S. A., Huang, A. C., Miazgowicz, M. M., Brassil, M. M., and Weinmann, A. S. (2008). Coordinated but physically separable interaction with H3K27-demethylase and H3K4-methyltransferase activities are required for T-box protein-mediated activation of developmental gene expression. Genes Dev. 22, 2980–2993. Mittnacht, S. (1998). Control of pRB phosphorylation. Curr. Opin. Genet Dev. 8, 21–27. Morris, E. J., and Dyson, N. J. (2001). Retinoblastoma protein partners. Adv. Cancer Res. 82, 1–54. Muller, C. W., and Herrmann, B. G. (1997). Crystallographic structure of the T domain-DNA complex of the Brachyury transcription factor. Nature 389, 884–888. Muller, H., Bracken, A. P., Vernell, R., Moroni, M. C., Christians, F., Grassilli, E., Prosperini, E., Vigo, E., Oliner, J. D., and Helin, K. (2001). E2Fs regulate the expression of genes involved in differentiation, development, proliferation, and apoptosis. Genes Dev. 15, 267–285. Naef, F., and Magnasco, M. O. (2003). Solving the riddle of the bright mismatches: labeling and effective binding in oligonucleotide arrays. Phys. Rev. E. Stat. Nonlin. Soft Matter Phys. 68, 011906 Nead, M. A., Baglia, L. A., Antinore, M. J., Ludlow, J. W., and McCance, D. J. (1998). Rb binds c-Jun and activates transcription. EMBO J. 17, 2342–2352. Nielsen, S. J., et al. (2001). Rb targets histone H3 methylation and HP1 to promoters. Nature 412, 561–565. Paxton, C., Zhao, H., Chin, Y., Langner, K., and Reecy, J. (2002). Murine Tbx2 contains domains that activate and repress gene transcription. Gene 283, 117–124. Polager, S., Kalma, Y., Berkovich, E., and Ginsberg, D. (2002). E2Fs upregulate expression of genes involved in DNA replication, DNA repair and mitosis. Oncogene 21, 437–446. Prince, S., Carreira, S., Vance, K. W., Abrahams, A., and Goding, C. R. (2004). Tbx2 directly represses the expression of the p21(WAF1) cyclin-dependent kinase inhibitor. Cancer Res. 64, 1669–1674. Pulichino, A. M., Vallette-Kasic, S., Tsai, J. P., Couture, C., Gauthier, Y., and Drouin, J. (2003). Tpit determines alternate fates during pituitary cell differentiation. Genes Dev. 17, 738–747. Ren, B., Cam, H., Takahashi, Y., Volkert, T., Terragni, J., Young, R. A., and Dynlacht, B. D. (2002). E2F integrates cell cycle progression with DNA repair, replication, and G(2)/M checkpoints. Genes Dev. 16, 245–256. Robertson, K. D., Ait-Si-Ali, S., Yokochi, T., Wade, P. A., Jones, P. L., and Wolffe, A. P. (2000). DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters. Nat. Genet 25, 338– 342. Rodriguez-Esteban, C., Tsukui, T., Yonei, S., Magallon, J., Tamura, K., and Izpisua Belmonte, J. C. (1999). The T-box genes Tbx4 and Tbx5 regulate limb outgrowth and identity. Nature 398, 814–818. Rodriguez, M., Aladowicz, E., Lanfrancone, L., and Goding, C. R. (2008). Tbx3 represses E-cadherin expression and enhances melanoma invasiveness. Cancer Res. 68, 7872–7881. Shih, H. H., Tevosian, S. G., and Yee, A. S. (1998). Regulation of differentiation by HBP1, a target of the retinoblastoma protein. Mol. Cell. Biol. 18, 4732–4743. Sinclair, C. S., et al. (2002). TBX2 is preferentially amplified in BRCA1- and BRCA2-related breast tumors. Cancer Res. 62, 3587–3591. Sinha, S., Abraham, S., Gronostajski, R. M., and Campbell, C. E. (2000). Differential DNA binding and transcription modulation by three T-box proteins, T, TBX1 and TBX2. Gene 258, 15–29. Suarez-Farinas, M., Pellegrino, M., Wittkowski, K. M., and Magnasco, M. O. (2005). Harshlight: a “corrective make-up” program for microarray chips. BMC Bioinform. 6, 294. Szabo, S. J., Kim, S. T., Costa, G. L., Zhang, X., Fathman, C. G., and Glimcher, L. H. (2000). A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell 100, 655–669. Takeuchi, J. K., Koshiba-Takeuchi, K., Matsumoto, K., Vogel-Hopker, A., Naitoh-Matsuo, M., Ogura, K., Takahashi, N., Yasuda, K., and Ogura, T. (1999). Tbx5 and Tbx4 genes determine the wing/leg identity of limb buds. Nature 398, 810–814. Tevosian, S. G., Shih, H. H., Mendelson, K. G., Sheppard, K. A., Paulson, K. E., and Yee, A. S. (1997). HBP1, a HMG box transcriptional repressor that is targeted by the retinoblastoma family. Genes Dev. 11, 383–396. Tong, Y., Aune, T., and Boothby, M. (2005). T-bet antagonizes mSin3a recruitment and transactivates a fully methylated IFN-gamma promoter via a conserved T-box half-site. Proc. Natl. Acad. Sci. USA 102, 2034–2039. Trouche, D., Le Chalony, C., Muchardt, C., Yaniv, M., and Kouzarides, T. (1997). RB and hbrm cooperate to repress the activation functions of E2F1. Proc. Natl. Acad. Sci. USA 94, 11268–11273. Vance, K. W., Carreira, S., Brosch, G., and Goding, C. R. (2005). Tbx2 is overexpressed and plays an important role in maintaining proliferation and suppression of senescence in melanomas. Cancer Res. 65, 2260–2268. Vormer, T. L., Foijer, F., Wielders, C. L., and te Riele, H. (2008). Anchorageindependent growth of pocket protein-deficient murine fibroblasts requires bypass of G2 arrest and can be accomplished by expression of TBX2. Mol. Cell. Biol. 28, 7263–7273. Yavuzer, U., Keenan, E., Lowings, P., Vachtenheim, J., Currie, G., and Goding, C. R. (1995). The Microphthalmia gene product interacts with the retinoblastoma protein in vitro and is a target for deregulation of melanocyte-specific transcription. Oncogene 10, 123–134. Yu, B. D., Becker-Hapak, M., Snyder, E. L., Vooijs, M., Denicourt, C., and Dowdy, S. F. (2003). Distinct and nonoverlapping roles for pRB and cyclin D:cyclin-dependent kinases 4/6 activity in melanocyte survival. Proc. Natl. Acad. Sci. USA 100, 14881–14886.
URI:	http://wrap.warwick.ac.uk/id/eprint/5409

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