Replication protein A confers structure-specific endonuclease activities to the XPF-ERCC1 and XPG subunits of human DNA repair excision nuclease

Tsukasa Matsunaga, Chi Hyun Park, Tadayoshi Bessho, David Mu, Aziz Sancar

Research output: Contribution to journalArticle

151 Citations (Scopus)

Abstract

XPF-ERCC1 and XPG proteins are nucleases that are involved in human nucleotide excision repair. In this study, we characterized the structure- specific junction-cutting activities of both nucleases using DNA substrates containing a bubble or loop structure. We found that the junction-cutting activities of XPF-ERCC1 and XPG were greatly stimulated by human replication protein A (RPA), while heterologous single-stranded DNA-binding proteins could not substitute for human RPA. To test for specific interaction between RPA and XPF-ERCC1 as is known to occur between RPA and XPG, we employed a pull-down assay with immobilized 'bubble' substrate. We found that the binding of XPF-ERCC1 complex to the bubble substrate was enhanced by RPA, suggesting a possible mechanism for RPA in the excision nuclease system, that is the targeting of the nuclease subunits to their specific sites of action. Furthermore, the RPA-promoted junction cutting by XPF-ERCC1 and XPG nucleases was observed with 'loop' substrates as well, raising the possibility that XPF-ERCC1, XPG, and RPA may function in removing loop structures from DNA, independent of the other subunits of the human excinuclease.

Original languageEnglish (US)
Pages (from-to)11047-11050
Number of pages4
JournalJournal of Biological Chemistry
Volume271
Issue number19
DOIs
StatePublished - Jun 4 1996

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Replication Protein A
Endonucleases
DNA Repair
Repair
DNA
Substrates
Deoxyribonucleases
DNA-Binding Proteins
Assays
Nucleotides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Replication protein A confers structure-specific endonuclease activities to the XPF-ERCC1 and XPG subunits of human DNA repair excision nuclease. / Matsunaga, Tsukasa; Park, Chi Hyun; Bessho, Tadayoshi; Mu, David; Sancar, Aziz.

In: Journal of Biological Chemistry, Vol. 271, No. 19, 04.06.1996, p. 11047-11050.

Research output: Contribution to journalArticle

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