Identification of USF2 as a key regulator of Runx2 expression in mouse pluripotent mesenchymal D1 cells.
Runx2 is one of the most important transcription factors directing the osteogenesis of mesenchymal stem cells and osteoblastic functions. It is likely that the factors controlling Runx2 expression would trigger the early steps of osteoblast differentiation. By using a reporter gene assay for 4.5 kb Runx2 promoter, it was found that the first 305 bp of Runx2 promoter are active in D1 cells. Within this region, electromobility shift assays (EMSAs) delineated a 6 bp of CACATG bound specifically by the proteins from D1 cell nuclear extract. Antibody super-shift and DNA-coupling magnetic bead pull-down assay indicated that the protein bound to this sequence is USF2. Site-specific mutagenesis revealed that this sequence contributed to the activity of 305 bp Runx2 promoter. Thus, we suggest that USF2 might be one of the regulators for the expression of the Runx2 gene in D1 cells.
Wang
Chihuei
C
Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan.
Lee
Grace
G
Hsu
Wayne
W
Yeh
Ching-Hua
CH
Ho
Mei-Ling
ML
Wang
Gwo-Jaw
GJ
D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D019281:Dimerization; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D051720:Upstream Stimulatory Factors / Q000378:metabolism*; D000099229:RE1-Silencing Transcription Factor
D050659:Core Binding Factor Alpha 1 Subunit; D004247:DNA; D012097:Repressor Proteins; D014157:Transcription Factors; D051720:Upstream Stimulatory Factors; D000099229:RE1-Silencing Transcription Factor; C493716:Runx2 protein, mouse; C497186:Usf2 protein, mouse