Multiple proteins are involved in the protein-DNA complex in the proximal promoter of the human alpha1(III) collagen gene (COL3A1).
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.
Yoshino
Tomoaki
T
Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
Sumiyoshi
Hideaki
H
Shin
Toshitaka
T
Matsuo
Noritaka
N
Inagaki
Yutaka
Y
Ninomiya
Yoshifumi
Y
Yoshioka
Hidekatsu
H
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
C439971:COL3A1 protein, human; D024061:Collagen Type III; D004268:DNA-Binding Proteins