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Volume 271, Number 45, Issue of November 8, 1996 pp. 28324-28330
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Isolation and Characterization of the Human Cytochrome P450 CYP1B1 Gene

(Received for publication, April 16, 1996, and in revised form, July 10, 1996)

Yong Ming Tang Dagger , Yu-Yuan P. Wo Dagger , Jane Stewart Dagger , Anita L. Hawkins § , Constance A. Griffin § , Thomas R. Sutter and William F. Greenlee par

From the Dagger  Department of Pharmacology and Toxicology, Purdue University, West Lafayette, Indiana 47907, the § Oncology Center and Departments of Pathology and Oncology and  Department of Environmental Health Sciences, Johns Hopkins Medical Institutions, Baltimore, Maryland 21205, and the par  Department of Pharmacology and Molecular Toxicology, University of Massachusetts Medical Center, Worcester, Massachusetts 01655

Previously, we identified a novel human cytochrome P450 cDNA that is inducible by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and represents the first member of a new subfamily designated cytochrome P4501B1 (CYP1B1; Sutter, T. R., Tang, Y. M., Hayes, C. L., Wo, Y. P., Jabs, E. W., Li, X., Yin, H., Cody, C. W., and Greenlee, W. F. (1994) J. Biol. Chem. 269, 13092-13099). Here, we report on the isolation and initial characterization of the CYP1B1 gene. The CYP1B1 gene maps to human chromosome 2 at 2p21-22 and contains three exons and two introns. The putative open reading frame starts in the second exon and is 1629 base pairs in length. Southern analysis using DNA probes directed to each of the three exons confirmed that CYP1B1 is a single copy gene. Human CYP1B1 differs from its two most closely related members of the cytochrome P450 superfamily, CYP1A1 and CYP1A2, in the number of exons (3 versus 7) and chromosome location (2 versus 15). A single transcription initiation site was identified by primer extension analysis and S1 nuclease mapping. Based on nucleotide sequence analysis, the CYP1B1 gene lacks a consensus TATA box in the promoter region and contains nine TCDD-responsive enhancer core binding motifs (5'-GCGTG-3') located within a 2.5-kilobase pair genomic fragment 5'-ward of the transcription initiation start site. Deletion analysis of chloramphenicol acetyltransferase reporter gene constructs containing 5' CYP1B1 genomic fragments indicates that a region from -1022 to -835 containing three of the nine core binding motifs contributes to the TCDD-inducible expression of CYP1B1.


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