Regulation of the bone-specific osteocalcin gene by p300 requires Runx2/Cbfa1 and the vitamin D3 receptor but not p300 intrinsic histone acetyltransferase activity

被引:156
作者
Sierra, J
Villagra, A
Paredes, R
Cruzat, F
Gutierrez, S
Javed, A
Arriagada, G
Olate, J
Imschenetzky, M
van Wijnen, AJ
Lian, JB
Stein, GS
Stein, JL
Montecino, M
机构
[1] Univ Concepcion, Fac Ciencias Biol, Dept Mol Biol, Concepcion, Chile
[2] Univ Massachusetts, Sch Med, Dept Cell Biol, Worcester, MA 01655 USA
关键词
D O I
10.1128/MCB.23.9.3339-3351.2003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
p300 is a multifunctional transcriptional coactivator that serves as an adapter for several transcription factors including nuclear steroid hormone receptors. p300 possesses an intrinsic histone acetyltransferase (HAT) activity that may be critical for promoting steroid-dependent transcriptional activation. In osteoblastic cells, transcription of the bone-specific osteocalcin (OC) gene is principally regulated by the Runx2/Cbfa1 transcription factor and is stimulated in response to vitamin D-3 via the vitamin D-3 receptor complex. Therefore, we addressed p300 control of basal and vitamin D-3-enhanced activity of the OC promoter. We find that transient overexpression of p300 results in a significant dose-dependent increase of both basal and vitamin D-3-stimulated OC gene activity. This stimulatory effect requires intact Runx2/Cbfa1 binding sites and the vitamin D-responsive element. In addition, by coimmunoprecipitation, we show that the endogenous Runx2/Cbfa1 and p300 proteins are components of the same complexes within osteoblastic cells under physiological concentrations. We also demonstrate by chromatin immunoprecipitation assays that p300, Runx2/Cbfa1, and 1alpha,25-dihydroxyvitamin D-3 receptor interact with the OC promoter in intact osteoblastic cells expressing this gene. The effect of p300 on the OC promoter is independent of its intrinsic HAT activity, as a HAT-deficient p300 mutant protein up-regulates expression and cooperates with P/CAF to the same extent as the wild-type p3OO. On the basis of these results, we propose that p300 interacts with key transcriptional regulators of the OC gene and bridges distal and proximal OC promoter sequences to facilitate responsiveness to vitamin D-3.
引用
收藏
页码:3339 / 3351
页数:13
相关论文
共 35 条
[1]   AP-1 and vitamin D receptor (VDR) signaling pathways converge at the rat osteocalcin VDR element:: Requirement for the internal activating protein-1 site for vitamin D-mediated trans-activation [J].
Aslam, F ;
McCabe, L ;
Frenkel, B ;
van Wijnen, AJ ;
Stein, GS ;
Lian, JB ;
Stein, JL .
ENDOCRINOLOGY, 1999, 140 (01) :63-70
[2]   An AML-1 consensus sequence binds an osteoblast-specific complex and transcriptionally activates the osteocalcin gene [J].
Banerjee, C ;
Hiebert, SW ;
Stein, JL ;
Lian, JB ;
Stein, GS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (10) :4968-4973
[3]  
BANERJEE U, 1997, LOOP TR RESTRUCT COM, V3, P1
[4]   IN-VIVO OCCUPANCY OF THE VITAMIN-D RESPONSIVE ELEMENT IN THE OSTEOCALCIN GENE SUPPORTS VITAMIN-D-DEPENDENT TRANSCRIPTIONAL UP-REGULATION IN INTACT-CELLS [J].
BREEN, EC ;
VANWIJNEN, AJ ;
LIAN, JB ;
STEIN, GS ;
STEIN, JL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (26) :12902-12906
[5]  
CHAKRAVARTI D, 1996, NATURE, V383, P9
[6]   Nuclear receptors: coactivators, corepressors and chromatin remodeling in the control of transcription [J].
Collingwood, TN ;
Urnov, FD ;
Wolffe, AP .
JOURNAL OF MOLECULAR ENDOCRINOLOGY, 1999, 23 (03) :255-275
[7]  
DUCY P, 1995, MOL CELL BIOL, V15, P1858
[8]   Osf2/Cbfa1: A transcriptional activator of osteoblast differentiation [J].
Ducy, P ;
Zhang, R ;
Geoffroy, V ;
Ridall, AL ;
Karsenty, G .
CELL, 1997, 89 (05) :747-754
[9]  
GOLDMAN R, 2000, GENE DEV, V14, P1553
[10]   CCAAT/enhancer-binding proteins (C/EBP) β and δ activate osteocalcin gene transcription and synergize with Runx2 at the C/EBP element to regulate bone-specific expression [J].
Gutierrez, S ;
Javed, A ;
Tennant, DK ;
van Rees, M ;
Montecino, M ;
Stein, GS ;
Stein, JL ;
Lian, JB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (02) :1316-1323