Nucleic Acids Research, Vol 25, Issue 16 3261-3268, Copyright © 1997 by Oxford University Press
SS Chen, EC Ruteshouser, SN Maity and B de Crombrugghe
We performed in vivo dimethylsulfate footprinting of the 220 bp mouse
proximal proalpha1(I) collagen promoter and the 350 bp mouse proximal
proalpha2(I) collagen promoter in BALB/3T3 fibroblasts, primary mouse skin
fibroblasts, S-194 B cells, NMuLi liver epithelial cells and RAG renal
adenocarcinoma cells and in vitro DNase I footprinting of these promoters
using nuclear extracts of these different cell types. Whereas proalpha1(I)
and proalpha2(I) collagen RNAs were present in BALB/3T3 fibroblasts and
primary fibroblasts, these RNAs could not be detected in the three other
cell lines. Comparison of in vitro DNase I footprints for each of the two
proximal collagen promoters indicated that the patterns of protection were
very similar with the different nuclear extracts, suggesting that the DNA
binding proteins binding to these promoters were present in all cell types
tested. In contrast, in vivo footprints over these proximal promoters were
cell-specific, occurring only in fibroblast cells and not in the other
three cell types. The in vivo footprints were generally located within the
in vitro footprinted regions. Our results suggest that although all cell
types tested contained nuclear proteins that can bind to the proximal
proalpha1(I) and proalpha2(I) collagen promoters in vitro , it is only in
fibroblasts that these proteins bind to their cognate sites in vivo . We
discuss possible regulatory mechanisms in type I collagen genes that can
contribute to the cell-specific in vivo protein-DNA interactions at the
proximal promoters.
ARTICLES
Cell-specific in vivo DNA-protein interactions at the proximal promoters of the pro alpha 1(I) and the pro alpha2(I) collagen genes
Department of Molecular Genetics, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX 77030, USA.
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