E2F1 and TFDP1 Regulate PITX1 Expression in Normal and Osteoarthritic Articular Chondrocytes

被引:22
作者
Pellicelli, Martin [1 ,2 ]
Picard, Cynthia [1 ,2 ]
Wang, DaShen [1 ]
Lavigne, Patrick [3 ,4 ]
Moreau, Alain [1 ,2 ,5 ]
机构
[1] St Justine Univ, Hosp Res Ctr, Viscogliosi Lab Mol Genet Musculoskeletal Dis, Montreal, PQ, Canada
[2] Univ Montreal, Dept Biochem & Mol Med, Fac Med, Montreal, PQ, Canada
[3] Maisonneuve Rosemont Hosp, Orthoped Div, Montreal, PQ, Canada
[4] Univ Montreal, Dept Surg, Fac Med, Montreal, PQ, Canada
[5] Univ Montreal, Dept Stomatol, Fac Dent, Montreal, PQ, Canada
来源
PLOS ONE | 2016年 / 11卷 / 11期
关键词
ENDOCHONDRAL OSSIFICATION; TRANSCRIPTION FACTORS; DNA-DAMAGE; APOPTOSIS; P53; ACCUMULATION; SENESCENCE; FAMILY; GENES; DP-1;
D O I
10.1371/journal.pone.0165951
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We previously reported a loss-of-PITX1 expression in patients suffering of knee/hip osteo-arthritis (OA). Search for the mechanism underlying this event led us to discover that PITX1 repression was triggered by the aberrant nuclear accumulation of Prohibitin (PHB1), an E2F1 co-repressor, in OA articular chondrocytes. In the current study, we assessed in details the involvement of E2F transcription factors in regulating PITX1 expression. We also analyzed other genes that are similarly regulated by E2F in regard to osteoarthritis. The transcriptional regulation of the PITX1 promoter by E2F1 was analyzed with the luciferase reporter assay, and chromatin immunoprecipitation assays, which confirmed direct E2F1-PITX1 interactions. The probable binding sites for E2F1 in the PITX1 promoter were identified by DNA pulldown experiments. In silico and in vitro analyses show that the PITX1 proximal promoter region contains 2 specific sequences that are bound by E2F1. Overexpression of E2F1 enhances PITX1 promoter activity and mRNA transcription. In primary control and osteoarthritis chondrocytes, real time RT-PCR was used to measure the mRNA expression levels of candidate genes under E2F1 transcriptional control. Transcription Factor Dp-1 (TFDP1) knockdown experiments confirmed that the E2F1-TFDP1 complex regulates PITX1. Knockdown of TFDP1, an E2F1 dimerization partner, inhibits the activating effect of E2F1 and reduces both PITX1 promoter activity and mRNA transcription. Real time RT-PCR results reveal reduced expression of TFDP1 and a similar downregulation of their targets PITX1, BRCA1, CDKN1A, and RAD51 in mid-stage OA chondrocytes. Collectively, our data define a previously uncharacterized role for E2F1 and TFDP1 in the transcriptional regulation of PITX1 in articular chondrocytes. Additional E2F1 targets may be affected in OA pathogenesis.
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页数:15
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