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

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Abstract

To determine the contribution of a human DNA repair gene, ERCC2 (XPD), to mutagenesis in 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 0.3% 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

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Xeroderma Pigmentosum
Genetic Vectors
Somatostatin-Secreting Cells
DNA Repair
Hypersensitivity
Genes
Plasmids
Mutagenesis
Survival
Mutation
Chloramphenicol O-Acetyltransferase
Transfection
Reference Values
Fibroblasts
Cell Line
Antibodies

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Gözükara, E. M., Parris, C. N., Weber, C. A., Salazar, E. P., Seidman, M. M., Watkins, J. F., ... Kraemer, K. H. (1994). The human DNA repair gene, ERCC2 (XPD), corrects ultraviolet hypersensitivity and ultraviolet hypermutability of a shuttle vector replicated in xeroderma pigmentosum group D cells. Cancer Research, 54(14), 3837-3844.

The human DNA repair gene, ERCC2 (XPD), corrects ultraviolet hypersensitivity and ultraviolet hypermutability of a shuttle vector replicated in xeroderma pigmentosum group D cells. / Gözükara, Engin M.; Parris, Christopher N.; Weber, Christine A.; Salazar, Edmund P.; Seidman, Michael M.; Watkins, John F.; Prakash, Louise; Kraemer, Kenneth H.

In: Cancer Research, Vol. 54, No. 14, 15.07.1994, p. 3837-3844.

Research output: Contribution to journalArticle

Gözükara, Engin M. ; Parris, Christopher N. ; Weber, Christine A. ; Salazar, Edmund P. ; Seidman, Michael M. ; Watkins, John F. ; Prakash, Louise ; Kraemer, Kenneth H. / The human DNA repair gene, ERCC2 (XPD), corrects ultraviolet hypersensitivity and ultraviolet hypermutability of a shuttle vector replicated in xeroderma pigmentosum group D cells. In: Cancer Research. 1994 ; Vol. 54, No. 14. pp. 3837-3844.
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abstract = "To determine the contribution of a human DNA repair gene, ERCC2 (XPD), to mutagenesis in 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 0.3{\%} 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.",
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AU - Parris, Christopher N.

AU - Weber, Christine A.

AU - Salazar, Edmund P.

AU - Seidman, Michael M.

AU - Watkins, John F.

AU - Prakash, Louise

AU - Kraemer, Kenneth H.

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