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Nucleic Acids Research, 1992, Vol. 20, No. 2 217-224
© 1992

MOLECULAR BIOLOGY

Molecular and functional analysis of the XPBC/ERCC-3 promoter: transcription activity is dependent on the integrity of an Sp1-binding site

Libin Ma, Geert Weeda1, Aart.G. Jochemsen, Dirk Bootsma1, Jan H.J. Hoeijmakers1 and Alex J. van der Eb*

Laboratory for Molecular Carcinogenesis, Medical Genetics Center, Sylvius Laboratories, University of Leiden Wassenaarseweg 72, 2333 AL Leiden 1Department of Cell Biology and Genetics, Medical Genetics Center, Erasmus University Rotterdam 3000 DR Rotterdam, The Netherlands

*To whom correspondence should be addressed

Received November 8, 1991. Accepted December 18, 1991.

The human XPBC/ERCC-3 gene, which corrects the excision-repair defect in xeroderma pigmentosum group B cells and the UV-sensitive CHO mutant 27–1 cells, appears to be expressed constitutively in various cell types and tissues. We have analysed the structure and functionality of the XPBC/ERCC-3 promoter. Transcription of the XPBC/ERCC-3 gene is initiated from heterogeneous sites, with a major startpoint mapped at position - 54 (relative to the translation start codon ATG). The promoter region does not possess classical TATA and CAAT elements, but it is GC-rich and contains three putative Sp1-binding sites. In addition, there are two elements related to the cyclic AMP (cAMP)-response element (CRE) and the 12–O-tetradecanoyl phorbol-13-acetate-response element (TRE) in the 5'-fIanking region. Transient expression analysis of XPBC/ERCC-3 promoter-CAT chimeric plasmids revealed that a 127-bp fragment, spanning position -129 to -3, is minimally required for the promoter activity. Transcription of the XPBC/ERCC-3 promoter depends on the integrity of a putative Sp1-binding site in close proximity to the major cap site. Band shift assays showed that this putative Sp1-binding site can interact specifically with a nuclear factor, most likely transcription factor Sp1 (or an Sp1-like factor) in vitro.


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