An interaction between the DNA repair factor XPA and replication protein A appears essential for nucleotide excision repair

Replication protein A (RPA) is required for simian virus 40-directed DNA replication in vitro and for nucleotide excision repair (NER). Here we report that RPA and the human repair protein XPA specifically interact both in vitro and in vivo. Mapping of the RPA-interactive domains in XPA revealed that both of the largest subunits of RPA, RPA-70 and RPA-34, interact with XPA at distinct sites. A domain involved in mediating the interaction with RPA-70 was located between XPA residues 153 and 176. Deletion of highly conserved motifs within this region identified two mutants that were deficient in binding RPA in vitro and highly defective in NER both in vitro and in vivo. A second domain mediating the interaction with RPA-34 was identified within the first 58 residues in XPA. Deletion of this region, however, only moderately affects the complementing activity of XPA in vivo. Finally, the XPA-RPA complex is shown to have a greater affinity for damaged DNA than XPA alone. Taken together, these results indicate that the interaction between XPA and RPA is required for NER but that only the interaction with RPA-70 is essential.

[1]  C. Ingles,et al.  RPA involvement in the damage-recognition and incision steps of nucleotide excision repair , 1995, Nature.

[2]  R. Legerski,et al.  Mutations in XPA that prevent association with ERCC1 are defective in nucleotide excision repair , 1995, Molecular and cellular biology.

[3]  A. Sancar,et al.  The General Transcription-Repair Factor TFIIH Is Recruited to the Excision Repair Complex by the XPA Protein Independent of the TFIIE Transcription Factor (*) , 1995, The Journal of Biological Chemistry.

[4]  A. Yasui,et al.  DNA Repair Protein XPA Binds Replication Protein A (RPA) (*) , 1995, The Journal of Biological Chemistry.

[5]  D. S. Hsu,et al.  Reconstitution of Human DNA Repair Excision Nuclease in a Highly Defined System (*) , 1995, The Journal of Biological Chemistry.

[6]  A. Sancar.  Mechanisms of DNA excision repair. , 1994, Science.

[7]  G. Lucchini,et al.  Replication factor A is required in vivo for DNA replication, repair, and recombination , 1994, Molecular and cellular biology.

[8]  A. Bardwell,et al.  Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease. , 1994, Science.

[9]  G. Hannon,et al.  The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA , 1994, Nature.

[10]  A. Sancar,et al.  Formation of a ternary complex by human XPA, ERCC1, and ERCC4(XPF) excision repair proteins. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Elledge,et al.  Specific association between the human DNA repair proteins XPA and ERCC1. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[12]  B. Stillman,et al.  Anatomy of a DNA replication fork revealed by reconstitution of SV40 DNA replication in vitro , 1994, Nature.

[13]  R. Wood,et al.  Preferential binding of the xeroderma pigmentosum group A complementing protein to damaged DNA. , 1993, Biochemistry.

[14]  J. Newport,et al.  Distinct roles of cdk2 and cdc2 in RP-A phosphorylation during the cell cycle. , 1993, Journal of cell science.

[15]  R. Wood,et al.  Co‐correction of the ERCC1, ERCC4 and xeroderma pigmentosum group F DNA repair defects in vitro. , 1993, The EMBO journal.

[16]  A. Yasui,et al.  Evidence for a repair enzyme complex involving ERCC1 and complementing activities of ERCC4, ERCC11 and xeroderma pigmentosum group F. , 1993, The EMBO journal.

[17]  J. Hoeijmakers,et al.  Nucleotide excision repair. II: From yeast to mammals. , 1993, Trends in genetics : TIG.

[18]  S. Elledge,et al.  The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. , 1993, Genes & development.

[19]  B. Stillman,et al.  An interaction between replication protein A and SV40 T antigen appears essential for primosome assembly during SV40 DNA replication. , 1993, The Journal of biological chemistry.

[20]  Xiaohua Wu,et al.  DNA repair synthesis during base excision repair in vitro is catalyzed by DNA polymerase epsilon and is influenced by DNA polymerases alpha and delta in Saccharomyces cerevisiae , 1993, Molecular and cellular biology.

[21]  R. Legerski,et al.  Expression cloning of a human DNA repair gene involved in xeroderma pigmentosum group C , 1992, Nature.

[22]  D. Lane,et al.  A role for the human single-stranded DNA binding protein HSSB/RPA in an early stage of nucleotide excision repair. , 1992, Nucleic acids research.

[23]  K. Tanaka,et al.  Mutational analysis of the structure and function of the xeroderma pigmentosum group A complementing protein. Identification of essential domains for nuclear localization and DNA excision repair. , 1992, The Journal of biological chemistry.

[24]  B. Stillman,et al.  cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication. , 1992, The EMBO journal.

[25]  R. Fotedar,et al.  Cell cycle regulated phosphorylation of RPA‐32 occurs within the replication initiation complex. , 1992, The EMBO journal.

[26]  A. Sancar,et al.  Purification of PCNA as a nucleotide excision repair protein. , 1992, Nucleic Acids Research.

[27]  R. Wood,et al.  Proliferating cell nuclear antigen is required for DNA excision repair , 1992, Cell.

[28]  T. Kelly,et al.  Interaction of DNA polymerase alpha‐primase with cellular replication protein A and SV40 T antigen. , 1992, The EMBO journal.

[29]  R. Legerski,et al.  Expression cloning of a human DNA repair gene involved in xeroderma pigmentosum group C , 1992, Nature.

[30]  K Tanaka,et al.  Molecular cloning of human XPAC gene homologs from chicken, Xenopus laevis and Drosophila melanogaster. , 1991, Biochemical and biophysical research communications.

[31]  T. Lindahl,et al.  Complementation of DNA repair in xeroderma pigmentosum group A cell extracts by a protein with affinity for damaged DNA. , 1991, The EMBO journal.

[32]  A. Kwong,et al.  Synthesis of DNA by DNA polymerase ε in vitro , 1991 .

[33]  R. Legerski,et al.  High-frequency transformation of human repair-deficient cell lines by an Epstein-Barr virus-based cDNA expression vector. , 1991, Gene.

[34]  S. Fields,et al.  The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[35]  W. Heyer,et al.  Characterization of a cDNA encoding the 70-kDa single-stranded DNA-binding subunit of human replication protein A and the role of the protein in DNA replication. , 1991, The Journal of biological chemistry.

[36]  T. Kelly,et al.  Effects of T antigen and replication protein A on the initiation of DNA synthesis by DNA polymerase alpha-primase , 1991, Molecular and cellular biology.

[37]  D. Lane,et al.  Requirement for the replication protein SSB in human DMA excision repair , 1991, Nature.

[38]  A. Kwong,et al.  Synthesis of DNA by DNA polymerase epsilon in vitro. , 1991, The Journal of biological chemistry.

[39]  A. Yasui,et al.  Analysis of a human DNA excision repair gene involved in group A xeroderma pigmentosum and containing a zinc-finger domain , 1990, Nature.

[40]  A. Kwong,et al.  The in vitro replication of DNA containing the SV40 origin. , 1990, The Journal of biological chemistry.

[41]  B. Stillman,et al.  Cell-cycle-regulated phosphorylation of DNA replication factor A from human and yeast cells. , 1990, Genes & development.

[42]  J. Hurwitz,et al.  The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication. , 1990, The Journal of biological chemistry.

[43]  S. Fields,et al.  A novel genetic system to detect protein–protein interactions , 1989, Nature.

[44]  P. Hough,et al.  ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication , 1989, Nature.

[45]  R. Wood,et al.  Complementation of the xeroderma pigmentosum DNA repair defect in cell-free extracts , 1988, Cell.

[46]  B. Stillman,et al.  Cellular factors required for multiple stages of SV40 DNA replication in vitro. , 1988, The EMBO journal.

[47]  T. Kelly,et al.  Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[48]  B. Stillman,et al.  Initiation of eukaryotic DNA replication in vitro. , 1988, BioEssays : news and reviews in molecular, cellular and developmental biology.

[49]  D. Lane,et al.  p53 and DNA polymerase α compete for binding to SV40 T antigen , 1987, Nature.

[50]  F. Dean,et al.  Replication of simian virus 40 origin-containing DNA in vitro with purified proteins. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[51]  S. Henikoff.  Unidirectional digestion with exonuclease III in DNA sequence analysis. , 1987, Methods in enzymology.

[52]  D. Lane,et al.  p53 and DNA polymerase alpha compete for binding to SV40 T antigen. , 1987, Nature.

[53]  R. Tjian,et al.  T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication , 1986, Molecular and cellular biology.

[54]  R. Legerski,et al.  A sensitive endonuclease probe for lesions in deoxyribonucleic acid helix structure produced by carcinogenic or mutagenic agents. , 1977, The Journal of biological chemistry.

[55]  Zhigang Wang,et al.  DNA Repair Synthesis during Base Excision Repair In Vitro Is Catalyzed by DNA Polymerase E and Is Influenced by DNA Polymerases a and 8 in Saccharomyces cerevisiae , 2022 .