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Interaction of the 4S polycyclic hydrocarbon-binding protein with the cytochrome P450c gene

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6060682
The 4S polycyclic hydrocarbon binding protein has been purified from rat liver and its properties examined. The protein was incubated with subclones from the P450c gene; it specifically interacted with a plasmid that contained the 5'-half of intron 1, exon 1 and 5'-flanking sequences. Exonuclease foot-printing after binding of the 4S protein to portions of the P450c gene showed protection at -200 and -400 bp from exon 1. The region -882 to +2545bp was constructed before a reporter, chloramphenicol acetyl transferase (CAT) gene in a plasmid that contained the SV40 ori, polyA signals, ampicillin resist gene. The P450c region contained promoter and putative regulatory sequences. The construct was transfected into rat hepatocytes, RL-PR-C and into rat hepatoma cells, H-4-11-E. After addition of 3-methylcholanthrene (3MC), CAT expression was induced. When the plasmid was constructed with the P450c fragment inverted, no CAT expression was seen. Deletion of -95 to -665 or from -238 to -660 bp eliminated the expression of CAT in response to 3MC. These experiments indicated the importance of this region in the induction of P450c by 3MC.
Research Organization:
Univ. of Nebraska Medical Center, Omaha
OSTI ID:
6060682
Report Number(s):
CONF-870644-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
Country of Publication:
United States
Language:
English