Template switching (TS) mediates damage bypass via a recombination-related mechanism involving PCNA polyubiquitination and polymerase δ-dependent DNA synthesis. Using two-dimensional gel electrophoresis and EM, here we characterize TS 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.

Visualization of recombination-mediated damage bypass by template switching / M. Giannattasio, K. Zwicky, C. Follonier, M. Foiani, M. Lopes, D. Branzei. - In: NATURE STRUCTURAL & MOLECULAR BIOLOGY. - ISSN 1545-9993. - 21:10(2014 Oct), pp. 884-892. [10.1038/nsmb.2888]

Visualization of recombination-mediated damage bypass by template switching

M. Giannattasio
Primo
;
M. Foiani;
2014

Abstract

Template switching (TS) mediates damage bypass via a recombination-related mechanism involving PCNA polyubiquitination and polymerase δ-dependent DNA synthesis. Using two-dimensional gel electrophoresis and EM, here we characterize TS 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.
English
Chromatids; DNA Polymerase III; DNA Replication; DNA, Cruciform; DNA, Fungal; Proliferating Cell Nuclear Antigen; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Ubiquitination; DNA Damage; DNA Repair; Recombination, Genetic; Templates, Genetic
Settore BIO/11 - Biologia Molecolare
Articolo
Esperti anonimi
Pubblicazione scientifica
ott-2014
21
10
884
892
9
Pubblicato
Periodico con rilevanza internazionale
pubmed
crossref
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info:eu-repo/semantics/article
Visualization of recombination-mediated damage bypass by template switching / M. Giannattasio, K. Zwicky, C. Follonier, M. Foiani, M. Lopes, D. Branzei. - In: NATURE STRUCTURAL & MOLECULAR BIOLOGY. - ISSN 1545-9993. - 21:10(2014 Oct), pp. 884-892. [10.1038/nsmb.2888]
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M. Giannattasio, K. Zwicky, C. Follonier, M. Foiani, M. Lopes, D. Branzei
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/257713
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