Regulation of the human SOX9 promoter by Sp1 and CREB.
The transcription factor SOX9 is essential for multiple steps during skeletal development, including mesenchymal cell chondrogenesis and endochondral bone formation. We recently reported that the human SOX9 proximal promoter region is regulated by the CCAAT-binding factor through two CCAAT boxes located within 100 bp of the transcriptional start site. Here we report that the human SOX9 proximal promoter is also regulated by the cyclic-AMP response element binding protein (CREB) and Sp1. We show by DNaseI protection and EMSA analysis that CREB and Sp1 interact with specific sites within the SOX9 proximal promoter region. By transient transfection analysis we also demonstrate that mutations of the CREB and Sp1 binding sites result in a profound reduction of SOX9 promoter activity. Chromatin immunoprecipitation (ChIP) assay demonstrated that both Sp1 and CREB interact with the SOX9 promoter in vivo. Finally, we demonstrate that IL-1beta treatment of chondrocytes isolated from human normal and osteoarthritic (OA) cartilage down-regulates SOX9 promoter activity, an effect accompanied by a reduction of Sp1 binding to the SOX9 proximal promoter.
Piera-Velazquez
Sonsoles
S
Department of Medicine, Division of Rheumatology, Thomas Jefferson University, Jefferson Medical College, 233 S. 10th Street, Room 509 BLSB, Philadelphia, PA 19107-5541, USA.
Hawkins
David F
DF
Whitecavage
Mary Kate
MK
Colter
David C
DC
Stokes
David G
DG
Jimenez
Sergio A
SA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D020219:Chondrogenesis; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D006609:High Mobility Group Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020218:Response Elements; D055758:SOX9 Transcription Factor; D016329:Sp1 Transcription Factor / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D017362:Cyclic AMP Response Element-Binding Protein; D006609:High Mobility Group Proteins; D053583:Interleukin-1beta; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C527933:Sox9 protein, mouse; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D003850:Deoxyribonuclease I