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(Received for publication, September 29,
1995; and in revised form, December 8, 1995) The mouse phenobarbital (PB)-inducible Cyp2b10 gene
promoter has been isolated and sequenced, and control of its expression
has been characterized. The 1405-base pair (bp) Cyp2b10 promoter sequence is 83% identical to the corresponding region
from the rat CYP2B2 gene. In addition to the lack of CA repeats,
differences include insertion of 42 base pairs (-123/-82
bp) into the middle of a consensus sequence to the so-called
``Barbie box.'' In this report, we have developed a primary
mouse hepatocyte culture system in which endogenous 2B10 mRNA as well
as Cyp2b10-driven CAT activity were induced by PB and
1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP), but not
by the 3-chloro derivative of TCPOBOP. Deletion analysis of the Cyp2b10 promoter identified a basal transcription element at
-64/-34 bp and a negative element at -971/-775
bp. Sequences contained within the -1404/-971 bp region are
responsible for the induced CAT activity. DNase I protection and gel
shift assays detected five major protein binding sites within the
-1404/-971 bp fragment, one of which shared high sequence
identity with a portion of a regulatory element in CYP2B2 gene
(Trottier, E., Belzil, A., Stoltz, C., and Anderson, A.(1995) Gene 158, 263-268). Our results indicate that sequences important
for PB-induced transcription of Cyp2b10 gene are located in
the distal promoter.
Volume 271,
Number 16,
Issue of April 19, 1996 pp. 9746-9753
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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