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(Received for publication, June 6, 1997, and in revised form, July 29, 1997)
From the Our laboratory has cloned the cDNA (Sutter,
T. R., Tang, Y. M., Hayes, C. L., Wo, Y.-Y. P.,
Jabs, E. W., Li, X., Yin, H., Cody, C. W., and Greenlee,
W. F. (1994) J. Biol. Chem. 269, 13092-13099) and gene (Tang, Y. M., Wo, Y.-Y. P., Jabs, E. W.,
Stewart, J. C., Sutter, T. R., and Greenlee, W. F. (1996) J. Biol. Chem. 271, 28324-28330) for human
CYP1B1, a new member of the cytochrome P450 superfamily.
Here, we report on the mapping and function of the CYP1B1
promoter. The CYP1B1 promoter is fully functional, when it
is uncoupled from upstream enhancer elements. Deletion analysis and
site-directed mutagenesis identified four regulatory elements required
for maximum promoter activity: two antisense Sp1 sites (
Volume 272, Number 42,
Issue of October 17, 1997
pp. 26702-26707
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
Functional Analysis of the Promoter for the Human
CYP1B1 Gene
,
and
Department of Molecular Pharmacology and
Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907 and the § Department of Pharmacology and Molecular
Toxicology, University of Massachusetts Medical Center,
Worcester, Massachusetts 01655-0126
84 to 89
and 68 to 73), a TATA-like box (34 to 29), and an initiator
motif (5 to +3). The initiator and the TATA-like elements are both
required for basal promoter activity, with enhanced activity mediated
by the two antisense Sp1 elements. The CYP1B1 initiator was
demonstrated by in vitro transcription analysis to be
a positioning element that maintained fidelity of transcription from a
single site. Specific binding to a CYP1B1 initiator probe
by human nuclear extract proteins was competed either by the highly
homologous murine terminal deoxynucleotidyl transferase initiator or,
to a lesser extent, by the adenovirus major late initiator. Taken
together, these results indicate that the structure and function of the
CYP1B1 promoter confers constitutive expression of the gene
and assures fidelity of transcription initiation from a single site.
The CYP1B1 promoter is distinct from the promoters of the
closely related cytochrome P450s CYP1A1 and
CYP1A2 and is structurally and functionally similar to the
promoters of constitutively expressed genes and at least two
viruses.
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