Characterization of the Human α1(VI) Collagen Promoter and its Comparison with Human α2(VI) Promoters

摘要: From a human cosmid library, we isolated clone (5B) with an insert of 32 kb, encoding the amino-terminal and 5′-end flanking region α1(VI) collagen gene. Exon 1 was found to be 194 bp contain 5′ untranslated plus 97 coding sequence. 2 consists 130bp, size that is conserved across chicken mouse species. S1-nuclease-protection assays primer-extension analysis, using mRNA from dermal fibroblasts, show presence multiple transcription start sites located in ≈20 nucleotides. Canonical TATA CAAT boxes, as α1 promoters, were absent promoter. The promoter positions–1 to–190, polypyrimidine/polypurine-rich containing 12 CCCTCCCC (CT element consensus) sequences has potential binding for Sp1 AP2 factors. These regulatory proteins bind α2(VI) promoters [Saitta, B. & Chu, M.-L. (1994) Eur. J. Biochem. 223, 675–682]. To test transcriptional activity promoter, transient transfection experiments DNA constructs performed fibroblasts fibrosarcoma (HT1080) cell lines. drive expression chloramphenicol acetyl transferase (CAT) results strong CAT at positions–1700,–298 and–257, while low positions–4400,–142 and–5 when transfected fibroblasts. also identified positive negative regions but did not identify them cells.