title: Reconstitution of recombination-associated DNA synthesis with human proteins.
abstract: The repair of DNA breaks by homologous recombination is a high-fidelity process, necessary for the maintenance of genome integrity. Thus, DNA synthesis associated with recombinational repair must be largely error-free. In this report, we show that human DNA polymerase delta (δ) is capable of robust DNA synthesis at RAD51-mediated recombination intermediates dependent on the processivity clamp PCNA. Translesion synthesis polymerase eta (η) also extends these substrates, albeit far less processively. The single-stranded DNA binding protein RPA facilitates recombination-mediated DNA synthesis by increasing the efficiency of primer utilization, preventing polymerase stalling at specific sequence contexts, and overcoming polymerase stalling caused by topological constraint allowing the transition to a migrating D-loop. Our results support a model whereby the high-fidelity replicative DNA polymerase δ performs recombination-associated DNA synthesis, with translesion synthesis polymerases providing a supportive role as in normal replication.
authors.affiliation: Department of Microbiology & Molecular Genetics, University of California, Davis, Davis, CA 95616-8665, USA.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Sneeden
authors.forename: Sara M
authors.initials: SM
authors.lastname: Grossi
authors.forename: Inger
authors.initials: I
authors.lastname: Tappin
authors.forename: Jerard
authors.initials: J
authors.lastname: Hurwitz
authors.forename: Wolf-Dietrich
authors.initials: WD
authors.lastname: Heyer
mesh_terms: D004247:DNA / Q000096:biosynthesis* / Q000378:metabolism; D004258:DNA Polymerase III / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051135:Rad51 Recombinase / Q000378:metabolism; D059767:Recombinational DNA Repair; D051716:Replication Protein A / Q000378:metabolism*; D000099270:Y-Family DNA Polymerases
chemical_list: D004247:DNA; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase; D018809:Proliferating Cell Nuclear Antigen; D051135:Rad51 Recombinase; D051716:Replication Protein A; C495153:RAD51 protein, human; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases
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