Reconstitution of recombination-associated DNA synthesis with human proteins

Jessica L. Sneeden, Sara M. Grossi, Inger Tappin, Jerard Hurwitz, Wolf Dietrich Heyer

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)4913-4925
Number of pages13
JournalNucleic Acids Research
Issue number9
StatePublished - May 2013

ASJC Scopus subject areas

  • Genetics


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