title: Multiple proteins are involved in the protein-DNA complex in the proximal promoter of the human alpha1(III) collagen gene (COL3A1).
abstract: We have characterized the proximal promoter of the human alpha1(III) collagen gene (COL3A1). Transient transfection assays using a series of chimeric constructs linked to the luciferase gene indicated that the segment from -96 to -34 is necessary to activate transcription. Electrophoretic mobility shift assays (EMSAs) showed that the multiple proteins form the DNA-protein complex in different combinations depending on the cell types. A competition assay using mutant oligonucleotides showed that the sequence 5'-GCTCTCATATTTCAGAA-3' (-79 to -63 bp) is critical for DNA-protein complex formation. This sequence is contained in the B element of mouse alpha1(III) collagen gene (Col3a1) reported by Ruteshouse and de Crombrugghe (J. Biol. Chem., 1993). In the rhabdomyosarcoma cell line, A204, at least two proteins of 92-118 kDa and 40-52 kDa are involved in the DNA-protein complex bound to this motif.
authors.affiliation: Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Yoshino
authors.forename: Hideaki
authors.initials: H
authors.lastname: Sumiyoshi
authors.forename: Toshitaka
authors.initials: T
authors.lastname: Shin
authors.forename: Noritaka
authors.initials: N
authors.lastname: Matsuo
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Inagaki
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Ninomiya
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Yoshioka
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D024061:Collagen Type III / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: C439971:COL3A1 protein, human; D024061:Collagen Type III; D004268:DNA-Binding Proteins