Runx2-mediated regulation of the zinc finger Osterix/Sp7 gene.
The zinc finger transcription factor Osterix (Osx) regulates bone formation and osteoblast differentiation in vitro and in vivo. We investigated the transcriptional mechanisms underlying the mouse Osx expression by isolating and characterizing its 5' upstream region. We performed 5' RACE on mRNA isolated from murine chondroprogenitor cells and determined a cap site of Osx approximately -99 nucleotides upstream of the initiation codon. Sequence analysis of this TATA-less promoter shows several putative response elements for Sox9, VDRE, Runx and Sp1. Transfection of the Osx promoter driving the luciferase reporter gene into C3H10T1/2 and ATDC5 cells shows a strong basal promoter activity between 565 bp and 2 kb. Deletion mutant analyses show that the most proximal 852 kb of the Osx promoter contains the highest activating domains, while strong repressive domains were identified between 1.8 and 2 kb. Over-expression experiments indicate that Runx2 significantly transactivates the Osx promoter by at least 2 fold indicating that Osx is downstream of Runx2 in mesenchymal cells. This up-regulation was abrogated when the Runx2 responsive element on the Osx promoter was mutated. Finally, we show that Runx2 specifically binds to this DNA element in the Osx promoter. Thus our results show for the first time Osx transcriptional regulation through the bone and cartilage related transcription factor Runx2.
Center for Musculoskeletal Research, Department of Orthopaedics, Box 665, University of Rochester Medical Center, Rochester, NY 14642, USA.
Yasuhiko
Y
Nishio
Yufeng
Y
Dong
Mark
M
Paris
Regis J
RJ
O'Keefe
Edward M
EM
Schwarz
Hicham
H
Drissi
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism*; D005091:Exons / Q000235:genetics; D006801:Humans; D007438:Introns / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D050436:Regulatory Elements, Transcriptional / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000235:genetics*; D015854:Up-Regulation / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
D050659:Core Binding Factor Alpha 1 Subunit; C493716:Runx2 protein, mouse; D000076182:Sp7 Transcription Factor; C479466:Sp7 protein, mouse; D014157:Transcription Factors