Mutational analysis of a function of xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair

Takehiro Kobayashi, Seiji Takeuchi, Masafumi Saijo, Yoshimichi Nakatsu, Hiroshi Morioka, Eiko Otsuka, Mitsuo Wakasugi, Osamu Nikaido, Kiyoji Tanaka

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Abstract

To analyze the function of the xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair, we examined repair of UV-induced cyclobutane pyrimidine dimer (CPD) in transcribed and non-transcribed strands of the dihydrofolate reductase gene of xeroderma pigmentosum group A (XP-A) cell line (XP12ROSV) which was transfected with various types of mutant XPA cDNA. The transfectant overexpressing mutant XPA with a defect in the interaction with either ERCC1, replication protein A (RPA), or general transcription factor TFIIH, showed more or loss decreased repair of CPD in each strand in parallel, while in the transfectant overexpressing R207G (Arg207 to Gly) mutant XPA derived from XP129, a UV-resistant XP12ROSV revertant, the rate of CPD repair was almost normal in each strand. We also examined the dose responses of the XPA protein on CPD repair in each strand by the modulation of the expression levels of wild-type or R207G mutant XPA using an inducible expression system, LacSwitch® promoter. There were good correlations between the rate of CPD repair in each strand and the amount of XPA protein produced in these Lac cells. Our results indicate that the XPA protein is equally important for the CPD repair in both transcribed and non-transcribed strands and that the R207G mutation found in XP129 may not be responsible for a selective defect in CPD repair in the non-transcribed strand in XP129.

Original languageEnglish
Pages (from-to)4662-4668
Number of pages7
JournalNucleic acids research
Volume26
Issue number20
DOIs
Publication statusPublished - Oct 15 1998
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Genetics

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