Transcriptional Regulation of Chondrogenesis by Coactivator Tip60 Via Chromatin Association with Sox9 and Sox5

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2008 Apr 9

PMID 18390577

Citations 46

Authors

Takako Hattori

Francoise Coustry

Shelley Stephens

Heidi Eberspaecher

Masaharu Takigawa

Hideyo Yasuda

Benoit de Crombrugghe

Affiliations

Soon will be listed here.

Abstract

Sox9 is a transcription factor of the SRY family required for several steps of chondrogenesis. It activates the expression of various chondrocyte-specific genes, but the mechanisms and role of cofactors involved in Sox9-regulated gene transcription are not fully understood. Here, we report on the characterization of a Tat interactive protein-60 (Tip60) as Sox9-associated protein identified in a yeast two-hybrid screen. Both in vitro and in vivo assays confirmed the specificity of interactions between Sox9 and Tip60 including the existence of an endogenous complex containing both polypeptides in chondrocytes. Gel shift assays showed the presence of a complex containing Sox9, Tip60 and the DNA of an enhancer region of the Col2a1 promoter. Reporter assays using a Col2a1 promoter with multimerized Col2a1 Sox9-binding sites indicated that Tip60 enhanced the transcriptional activity of Sox9. A larger Col2a1 promoter showed that Tip60 increased the activity of this promoter in the presence of both Sox9 and Sox5. Ectopic expression of Sox9 and transient-cotransfection with Tip60 in COS7 cells showed a more diffuse subnuclear colocalization, suggesting changes in the chromatin structure. Chromatin immunoprecipitation assays showed that Tip60, Sox9 and Sox5 associated with the same Col2a1 enhancer region. Consistent with a role of Tip60 in chondrogenesis, addition of Tip60 siRNA to limb-bud micromass cultures delayed chondrocyte differention. Tip60 enhances acetylation of Sox9 mainly through K61, 253, 398 residues; however, the K61/253/398A mutant of Sox9 still exhibited enhanced transcriptional activity by Tip60. Our results support the hypothesis that Tip60 is a coactivator of Sox9 in chondrocytes.

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References

Smits P, Li P, Mandel J, Zhang Z, Deng J, Behringer R . The transcription factors L-Sox5 and Sox6 are essential for cartilage formation. Dev Cell. 2001; 1(2):277-90. DOI: 10.1016/s1534-5807(01)00003-x. View

Solursh M . Chondrogenesis: in vitro approaches. Pathol Immunopathol Res. 1988; 7(1-2):48-50. DOI: 10.1159/000157092. View

Sudbeck P, Schmitz M, Baeuerle P, Scherer G . Sex reversal by loss of the C-terminal transactivation domain of human SOX9. Nat Genet. 1996; 13(2):230-2. DOI: 10.1038/ng0696-230. View

Bi W, Huang W, Whitworth D, Deng J, Zhang Z, Behringer R . Haploinsufficiency of Sox9 results in defective cartilage primordia and premature skeletal mineralization. Proc Natl Acad Sci U S A. 2001; 98(12):6698-703. PMC: 34415. DOI: 10.1073/pnas.111092198. View

Kim M, Woo E, Chong Y, Homenko D, Kraus W . Acetylation of estrogen receptor alpha by p300 at lysines 266 and 268 enhances the deoxyribonucleic acid binding and transactivation activities of the receptor. Mol Endocrinol. 2006; 20(7):1479-93. PMC: 1483068. DOI: 10.1210/me.2005-0531. View

Fechner P . The role of SRY in mammalian sex determination. Acta Paediatr Jpn. 1996; 38(4):380-9. DOI: 10.1111/j.1442-200x.1996.tb03512.x. View

Lefebvre V, Huang W, Harley V, Goodfellow P, de Crombrugghe B . SOX9 is a potent activator of the chondrocyte-specific enhancer of the pro alpha1(II) collagen gene. Mol Cell Biol. 1997; 17(4):2336-46. PMC: 232082. DOI: 10.1128/MCB.17.4.2336. View

Hattori T, Eberspaecher H, Lu J, Zhang R, Nishida T, Kahyo T . Interactions between PIAS proteins and SOX9 result in an increase in the cellular concentrations of SOX9. J Biol Chem. 2006; 281(20):14417-28. DOI: 10.1074/jbc.M511330200. View

Legube G, Linares L, Tyteca S, Caron C, Scheffner M, Chevillard-Briet M . Role of the histone acetyl transferase Tip60 in the p53 pathway. J Biol Chem. 2004; 279(43):44825-33. DOI: 10.1074/jbc.M407478200. View

10.

Sharma M, Zarnegar M, Li X, Lim B, Sun Z . Androgen receptor interacts with a novel MYST protein, HBO1. J Biol Chem. 2000; 275(45):35200-8. DOI: 10.1074/jbc.M004838200. View

11.

Li B, Carey M, Workman J . The role of chromatin during transcription. Cell. 2007; 128(4):707-19. DOI: 10.1016/j.cell.2007.01.015. View

12.

Koopman P, Gubbay J, Vivian N, Goodfellow P, Lovell-Badge R . Male development of chromosomally female mice transgenic for Sry. Nature. 1991; 351(6322):117-21. DOI: 10.1038/351117a0. View

13.

Blaise R, GROBER J, Rouet P, Tavernier G, Daegelen D, Langin D . Testis expression of hormone-sensitive lipase is conferred by a specific promoter that contains four regions binding testicular nuclear proteins. J Biol Chem. 1999; 274(14):9327-34. DOI: 10.1074/jbc.274.14.9327. View

14.

Lefebvre V, Behringer R, de Crombrugghe B . L-Sox5, Sox6 and Sox9 control essential steps of the chondrocyte differentiation pathway. Osteoarthritis Cartilage. 2001; 9 Suppl A:S69-75. DOI: 10.1053/joca.2001.0447. View

15.

Zhou G, Garofalo S, Mukhopadhyay K, Lefebvre V, Smith C, Eberspaecher H . A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. J Cell Sci. 1995; 108 ( Pt 12):3677-84. DOI: 10.1242/jcs.108.12.3677. View

16.

Owens E, Solursh M . In vitro histogenic capacities of limb mesenchyme from various stage mouse embryos. Dev Biol. 1981; 88(2):297-311. DOI: 10.1016/0012-1606(81)90173-1. View

17.

Yang X . The diverse superfamily of lysine acetyltransferases and their roles in leukemia and other diseases. Nucleic Acids Res. 2004; 32(3):959-76. PMC: 384351. DOI: 10.1093/nar/gkh252. View

18.

Georgiakaki M, Chabbert-Buffet N, Dasen B, Meduri G, Wenk S, Rajhi L . Ligand-controlled interaction of histone acetyltransferase binding to ORC-1 (HBO1) with the N-terminal transactivating domain of progesterone receptor induces steroid receptor coactivator 1-dependent coactivation of transcription. Mol Endocrinol. 2006; 20(9):2122-40. DOI: 10.1210/me.2005-0149. View

19.

Vogel A, Tickle C . FGF-4 maintains polarizing activity of posterior limb bud cells in vivo and in vitro. Development. 1993; 119(1):199-206. DOI: 10.1242/dev.119.1.199. View

20.

Sterner D, Berger S . Acetylation of histones and transcription-related factors. Microbiol Mol Biol Rev. 2000; 64(2):435-59. PMC: 98999. DOI: 10.1128/MMBR.64.2.435-459.2000. View