The Human DNA Repair Gene, ERCC2 (XPD), Corrects Ultraviolet Hypersensitivity and Ultraviolet Hypermutability of a Shuttle Vector Replicated in Xeroderma Pigmentosum Group D Cells

Engin M. Gözükara, Christopher N. Parris, Christine A. Weber, Edmund P. Salazar, Michael M. Seidman, John F. Watkins, Louise Prakash, Kenneth H. Kraemer

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

To determine the contribution of a human DNA repair gene, ERCC2 (XPD), to mutagenesis n human cells, two ERCC2 (XPD)-transformed xeroderma pigmentosum complementation group D (XPD) cell lines with increased UV survival compared to XP6BE(SV40), the original XPD line, were studied: D6BE-ER2-2 with slightly increased UV survival; and D6BE-ER2-9 with normal UV survival. ERCC2 (XPD) antibody-reactive protein levels were elevated 4.8-fold in D6BE-ER2-2 and 17.6-fold in D6BE-ER2-9 relative to XP6BE(SV40). DNA repair ability was assessed by measuring the ability of the cells to restore expression to UV-treated plasmids. Transfection of pRSVcat exposed to 1000 J/m2 UV resulted in 03% chloramphenicol acetyltransferase activity in XP6BE(SV40) cells but 20-80% in D6BE-ER2-2, D6BE-ER2-9, and repair-proficient cells compared to untreated control plasmids. The UV hypersensitivity of the mutagenesis shuttle vector pSP189 in XP6BE(SV40) cells was partially corrected and the UV hypermutability and excess of G:C- A:T mutations of pSP189 fell to the normal range in D6BE-ER2-2 and D6BE-ER2-9 cells. However, the frequency of plasmids recovered with multiple base substitution mutations was significantly reduced with XP6BE(SV40) cells and remained low in D6BE-ER2-2 and D6BE-ER2-9 cells, when compared with the normal fibroblasts. The human DNA excision repair gene, ERCC2 (XPD), substantially corrected the plasmid UV hypersensitivity and UV hypermutability of xeroderma pigmentosum complementation group D cells; however, the dose response relationship varied for different end points.

Original languageEnglish (US)
Pages (from-to)3837-3844
Number of pages8
JournalCancer Research
Volume54
Issue number14
StatePublished - Jul 15 1994

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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