Interaction between GC box binding factors and Smad proteins modulates cell lineage-specific alpha 2(I) collagen gene transcription.
Type I collagen is produced predominantly in mesenchymal cells, but molecular mechanisms responsible for cell type-specific expression are virtually unknown. During fibrogenic process in the liver, activated hepatic stellate cells (HSC) are the main producers of type I collagen, whereas parenchymal hepatocytes produce little, if any, of this protein. We have previously reported that Sp1 and an interacting unknown factor(s) bind to the -313 to -255 sequence of the alpha2(I) collagen gene (COL1A2) and play essential roles for basal and TGF-beta-stimulated transcription in skin fibroblasts and HSC. Recently, Smad3 has been shown to bind to this region, and its interaction with Sp1 has been implicated in TGF-beta-elicited COL1A2 stimulation. The present study demonstrates predominant binding of Sp3 rather than Sp1 to this regulatory element in parenchymal hepatocytes. In these cells, this region did not exhibit strong enhancer activity or mediate the effect of TGF-beta. Transfection of HSC with an Sp3 expression plasmid abolished the COL1A2 response to TGF-beta, whereas overexpression of Sp1 in hepatocytes increased basal COL1A2 transcription and conferred TGF-beta responsiveness. Functional and physical interactions between Sp1 and Smad3, but not between Sp3 and Smad3, were demonstrated using the bacterial GAL4 system and immunoprecipitation-Western blot analyses. These results indicate that cell lineage-specific interactions between GC box binding factors and Smad protein(s) may account, at least in part, for differential COL1A2 transcription and TGF-beta responsiveness in HSC and parenchymal hepatocytes.
Department of Internal Medicine and Division of Clinical Research, National Kanazawa Hospital, Kanazawa 920-8650, Japan.
Y
Y
Inagaki
T
T
Nemoto
A
A
Nakao
Dijke Pt
K
K
Kobayashi
K
K
Takehara
P
P
Greenwel
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics*; D024042:Collagen Type I; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012867:Skin / Q000378:metabolism*; D051900:Smad3 Protein; D016329:Sp1 Transcription Factor / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D024042:Collagen Type I; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D051900:Smad3 Protein; C497787:Smad3 protein, rat; D016329:Sp1 Transcription Factor; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D003094:Collagen