Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY
Volume 21, Issue 10, Pages 884-892Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nsmb.2888
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Funding
- European Research Council (ERG)
- Associazione Italiana per is Ricerca sul Cancro (AIRC)
- Fondazione Telethon grants
- Swiss National Science Foundation [PP00P3_135292, 31003A_146924]
- AIRC grant
- Swiss National Science Foundation (SNF) [PP00P3_135292, 31003A_146924] Funding Source: Swiss National Science Foundation (SNF)
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Template switching (TS) mediates damage bypass via a recombination-related mechanism involving PCNA polyubiquitination and polymerase delta-dependent DNA synthesis. Using two-dimensional gel electrophoresis and EM, here we characterize IS intermediates arising in Saccharomyces cerevisiae at a defined chromosome locus, identifying five major families of intermediates. Single-stranded DNA gaps of 150-200 nt, and not DNA ends, initiate TS by strand invasion. This causes reannealing of the parental strands and exposure of the nondamaged newly synthesized chromatid, which serves as a replication template for the other blocked nascent strand. Structures resembling double Holliday junctions, postulated to be central double-strand break-repair intermediates but so far visualized only in meiosis, mediate late stages of TS before being processed to hemicatenanes. Our results reveal the DNA transitions accounting for recombination-mediated DNA-damage tolerance in mitotic cells and replication under conditions of genotoxic stress.
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