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PMID: 7565690 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 15 ·No. 10 ·1995-10-00 ·Pages 5396-402

Li L, Lu X, Peterson CA, Legerski RJ

Abstract

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.

MeSH Terms
Amino Acid Sequence Base Sequence Conserved Sequence DNA/metabolism DNA Damage DNA Repair/physiology DNA-Binding Proteins/genetics,metabolism Humans Molecular Sequence Data Recombinant Fusion Proteins/biosynthesis Replication Protein A Sequence Deletion Xeroderma Pigmentosum Group A Protein
Chemicals
DNA-Binding Proteins RPA1 protein, human Recombinant Fusion Proteins Replication Protein A XPA protein, human Xeroderma Pigmentosum Group A Protein DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Li L
Department of Molecular Genetics, University of Texas M. D. Anderson Cancer Center, Houston 77030, USA.
Lu X
Peterson C A
Legerski R J
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.10.5396
Published
1995-10-00
Pages
5396-402
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230789
Subset
IM
Grants
NCI NIH HHS · CA-52461 · United States
External Links
PubMed source DOI: 10.1128/MCB.15.10.5396 PMC Full Text View journal details
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