Human xeroderma pigmentosum group A protein interacts with human replication protein A and inhibits DNA replication.
Human replication protein A (RPA; also known as human single-stranded DNA binding protein, or HSSB) is a multisubunit complex involved in both DNA replication and repair. While the role of RPA in replication has been well studied, its function in repair is less clear, although it is known to be involved in the early stages of the repair process. We found that RPA interacts with xeroderma pigmentosum group A complementing protein (XPAC), a protein that specifically recognizes UV-damaged DNA. We examined the effect of this XPAC-RPA interaction on in vitro simian virus 40 (SV40) DNA replication catalyzed by the monopolymerase system. XPAC inhibited SV40 DNA replication in vitro, and this inhibition was reversed by the addition of RPA but not by the addition of DNA polymerase alpha-primase complex, SV40 large tumor antigen, or topoisomerase I. This inhibition did not result from an interaction between XPAC and single-stranded DNA (ssDNA), or from competition between RPA and XPAC for DNA binding, because XPAC does not show any ssDNA binding activity and, in fact, stimulates RPA's ssDNA binding activity. Furthermore, XPAC inhibited DNA polymerase alpha activity in the presence of RPA but not in RPA's absence. These results suggest that the inhibitory effect of XPAC on DNA replication probably occurs through its interaction with RPA.
Lee
S H
SH
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101-0318, USA.
Kim
D K
DK
Drissi
R
R
D004265:DNA Helicases / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D019384:Nucleic Acid Synthesis Inhibitors; D051716:Replication Protein A; D013539:Simian virus 40 / Q000235:genetics*; D014983:Xeroderma Pigmentosum / Q000235:genetics / Q000378:metabolism*; D051760:Xeroderma Pigmentosum Group A Protein
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D019384:Nucleic Acid Synthesis Inhibitors; C497167:RPA1 protein, human; D051716:Replication Protein A; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases