Accurate repair of non‐cohesive, double strand breaks in <i>Saccharomyces cerevisiae</i>: enhancement by homology‐assisted end‐joining

Moscariello, Mario; Florio, Carolina; Pulitzer, John F.

doi:10.1002/yea.1789

Cited by 8 publications

(4 citation statements)

References 34 publications

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“…mutant has a reduced repair efficiency compared to wild type, confirming our previous results obtained with plasmid pRS315. However, the yku70Δ mutant exhibits a lower repair efficiency than the sub1Δ mutant (P<0.05), indicating that the DNA sequence flanking the DNA break may affect the efficiency of NHEJ [50]. Nonetheless, the sub1Δ yku70Δ double mutant does not exhibit an additive deficiency, suggesting Sub1's role in NHEJ depends on Yku70.…”

Section: Resultsmentioning

confidence: 97%

Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair

Volkert

2013

PLoS ONE

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Non homologous end joining (NHEJ) is an important process that repairs double strand DNA breaks (DSBs) in eukaryotic cells. Cells defective in NHEJ are unable to join chromosomal breaks. Two different NHEJ assays are typically used to determine the efficiency of NHEJ. One requires NHEJ of linearized plasmid DNA transformed into the test organism; the other requires NHEJ of a single chromosomal break induced either by HO endonuclease or the I-SceI restriction enzyme. These two assays are generally considered equivalent and rely on the same set of NHEJ genes. PC4 is an abundant DNA binding protein that has been suggested to stimulate NHEJ. Here we tested the role of PC4's yeast homolog SUB1 in repair of DNA double strand breaks using different assays. We found SUB1 is required for NHEJ repair of DSBs in plasmid DNA, but not in chromosomal DNA. Our results suggest that these two assays, while similar are not equivalent and that repair of plasmid DNA requires additional factor(s) that are not required for NHEJ repair of chromosomal double-strand DNA breaks. Possible roles for Sub1 proteins in NHEJ of plasmid DNA are discussed.

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Section: Resultsmentioning

confidence: 97%

Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair

Volkert

2013

PLoS ONE

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“…In contrast, extended deletions are recovered in yku70 mutant strains [19], [56], [57]. An alternative end-joining pathway (MMEJ), which increases upon Ku loss, has also been described in a chromosomal context [7].…”

Section: C-nhej Is a Conservative But Versatile Dsb Repair Processmentioning

confidence: 99%

Is Non-Homologous End-Joining Really an Inherently Error-Prone Process?

2014

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DNA double-strand breaks (DSBs) are harmful lesions leading to genomic instability or diversity. Non-homologous end-joining (NHEJ) is a prominent DSB repair pathway, which has long been considered to be error-prone. However, recent data have pointed to the intrinsic precision of NHEJ. Three reasons can account for the apparent fallibility of NHEJ: 1) the existence of a highly error-prone alternative end-joining process; 2) the adaptability of canonical C-NHEJ (Ku- and Xrcc4/ligase IV–dependent) to imperfect complementary ends; and 3) the requirement to first process chemically incompatible DNA ends that cannot be ligated directly. Thus, C-NHEJ is conservative but adaptable, and the accuracy of the repair is dictated by the structure of the DNA ends rather than by the C-NHEJ machinery. We present data from different organisms that describe the conservative/versatile properties of C-NHEJ. The advantages of the adaptability/versatility of C-NHEJ are discussed for the development of the immune repertoire and the resistance to ionizing radiation, especially at low doses, and for targeted genome manipulation.

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“…Deletion of RSC subunits results in diminished precise and imprecise NHEJ frequency at the site of an HO induced chromosomal DSB (Shim et al 2005). Plasmid repair assays also revealed NHEJ defects upon deletion of RSC subunits (Florio et al 2007; Moscariello et al 2010; Shim et al 2005), although one group detected increased plasmid repair in rsc1Δ and rsc2Δ strains (Chai et al 2005). The RSC subunit Sth1 is detected at an HO endonuclease-induced DSB by ChIP as early as 10 minutes after break induction (Shim et al 2007; Shim et al 2005).…”

Section: Nhej and Chromatinmentioning

confidence: 99%

Consider the workhorse: Nonhomologous end-joining in budding yeast

Emerson

Bertuch

2016

Biochem. Cell Biol.

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DNA double strand breaks (DSBs) are dangerous sources of genome instability and must be repaired by the cell. Nonhomologous end joining (NHEJ) is an evolutionarily conserved pathway to repair DSBs by direct ligation of the ends, with no requirement for a homologous template. While NHEJ is the primary DSB repair pathway in mammalian cells, conservation of the core NHEJ factors throughout eukaryotes make the pathway attractive for study in model organisms. The budding yeast, Saccharomyces cerevisiae, has been used extensively to develop a functional picture of NHEJ. In this review, we will discuss the current understanding of NHEJ in S. cerevisiae. Topics include canonical end-joining, alternative end-joining, and pathway regulation. Particular attention will be paid to the NHEJ mechanism involving core factors, including Yku70/80, Dnl4, Lif1, and Nej1, as well as the various factors implicated in the processing of the broken ends. The relevance of chromatin dynamics to NHEJ will also be discussed. This review illustrates the use of S. cerevisiae as a powerful system to understand the principles of NHEJ, as well as in pioneering the direction of the field.

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Accurate repair of non‐cohesive, double strand breaks in Saccharomyces cerevisiae: enhancement by homology‐assisted end‐joining

Cited by 8 publications

References 34 publications

Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair

Differential Requirement for SUB1 in Chromosomal and Plasmid Double-Strand DNA Break Repair

Is Non-Homologous End-Joining Really an Inherently Error-Prone Process?

Consider the workhorse: Nonhomologous end-joining in budding yeast

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