Insulin-like growth factor-I regulates transcription of the elastin gene through a putative retinoblastoma control element. A role for Sp3 acting as a repressor of elastin gene transcription.
Previous studies have demonstrated that insulin-like growth factor-I (IGF-I) increases elastin gene transcription in aortic smooth muscle cells and that this up-regulation is accompanied by a loss of protein binding to the proximal promoter. Sp1 has been identified as one of the factors whose binding is lost, and in the present study we show that Sp3 binding is also abrogated by IGF-I, but in a selected manner. In functional analyses using Drosophila SL-2 cells, Sp1 expression can drive transcription from the elastin proximal promoter, while co-expression of Sp3 results in a repression of Sp1 activity. Footprint and gel shift analyses position the IGF-I responsive sequences to a putative retinoblastoma control element (RCE). Mutation of the putative RCE sequence as assessed by transient transfection of smooth muscle cells results in an increase in reporter activity equal in magnitude to that conferred by IGF-I on the wild type promoter. Together these results support the hypothesis that IGF-I-mediated increase in elastin transcription occurs via a mechanism of derepression involving the abrogation of a repressor that appears to be Sp3 binding to the RCE.
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA. foster@med-biochem.bu.edu
K J
KJ
Conn
C B
CB
Rich
D E
DE
Jensen
M R
MR
Fontanilla
M M
MM
Bashir
J
J
Rosenbloom
J A
JA
Foster
D000818:Animals; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000502:physiology*; D004331:Drosophila melanogaster; D004549:Elastin / Q000235:genetics*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D016296:Mutagenesis; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D012175:Retinoblastoma / Q000235:genetics*; D051705:Sp3 Transcription Factor; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*; D014162:Transfection
D004268:DNA-Binding Proteins; C497121:SP3 protein, human; C497123:Sp3 protein, rat; D014157:Transcription Factors; D051705:Sp3 Transcription Factor; D007334:Insulin-Like Growth Factor I; D004549:Elastin