PMID:8626644
| Citation |
Matsunaga, T, Park, CH, Bessho, T, Mu, D and Sancar, A (1996) Replication protein A confers structure-specific endonuclease activities to the XPF-ERCC1 and XPG subunits of human DNA repair excision nuclease. J. Biol. Chem. 271:11047-50 |
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| 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. |
| Links | |
| Keywords |
Base Sequence; Cloning, Molecular; DNA Repair; DNA Replication; DNA-(Apurinic or Apyrimidinic Site) Lyase; DNA-Binding Proteins/biosynthesis; DNA-Binding Proteins/isolation & purification; DNA-Binding Proteins/metabolism; Deoxyribonuclease IV (Phage T4-Induced); Endonucleases/metabolism; Humans; Kinetics; Lyases/metabolism; Molecular Sequence Data; Nuclear Proteins; Nucleic Acid Conformation; Oligodeoxyribonucleotides; Protein Biosynthesis; Proteins/isolation & purification; Proteins/metabolism; Recombinant Proteins/biosynthesis; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Replication Protein A; Sequence Tagged Sites; Substrate Specificity; Transcription Factors |
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