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Cloning and characterization of the promoter of baboonXRCC1, a gene involved in DNA strand-break repair

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Somatic Cell and Molecular Genetics

Abstract

The DNA repair geneXRCC1 was the first cloned human DNA repair gene involved in resistance to ionizing radiation. Previous studies have shown that rodent and baboon homologs ofXRCC1 are expressed in all tested tissues with significantly higher levels in testis. Furthermore, expression of murineXRCC1 is most abundant in pachytene spermatocytes and round spermatids. To begin to study regulation ofXRCC1 expression, the 5′ region of baboonXRCC1 was cloned and characterized. 400 bp of 5′-flanking region showed the greatest promoter activity, while −194 to −8 bp of the 5′-flanking region displayed core promoter activity in transient transfection assays. A comparison between baboon and human 5′-flanking sequences in the core promoter region revealed a potential CAAT-box, an imperfect CREB-binding site and two putative Sp1-binding sites. Results from transient transfection assays in which each putative binding site was individually mutated, indicated that the distal Sp1-binding site has a functional role in transcription. In comparison, both putative Sp1-binding sites bound protein(s) from HeLa cell nuclear extractsin vitro. In vitro binding was lost when mutated Sp1 sites were used in gel mobility shift assays. Finally, anti-Sp1 antibodies produced mobility supershifts, thereby indicating Sp1 or an Sp1-like protein bound to the DNA fragmentin vitro.

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  1. Department of Cellular and Structural Biology, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, 78240-7762, San Antonio, Texas

    Zi-Qiang Zhou & Christi A. Walter

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  1. Zi-Qiang Zhou
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Zhou, ZQ., Walter, C.A. Cloning and characterization of the promoter of baboonXRCC1, a gene involved in DNA strand-break repair. Somat Cell Mol Genet 24, 23–39 (1998). https://doi.org/10.1007/BF02677493

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  • DOI: https://doi.org/10.1007/BF02677493

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