A proteome-wide visual screen identifies fission yeast proteins localizing to DNA double-strand breaks

Yu, Yang; Ren, Jingyi; Zhang, Jiamin; Suo, Fang; Fang, Xiuqi; Wu, Fan; Du, Li-Lin

doi:10.1016/j.dnarep.2013.04.001

Cited by 37 publications

(49 citation statements)

References 67 publications

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“…We identified one protein, Ygr042w, with no known role in recombination repair. Mutants in YGR042W affect telomere length (Askree et al 2004), and the fission yeast homolog of Ygr042w, Dbl2, forms foci that colocalize with an induced double-strand DNA break (Yu et al 2013). The extensive colocalization of Ygr042w with Rad52 foci, similar to the extent of colocalization observed for members of the Rad52 epistasis group (Symington 2002) Rad55, Rad57, and Rad59, suggests that Ygr042w could function in repair of double-strand DNA breaks.…”

Section: Resultssupporting

confidence: 55%

“…Proteins that localize in nuclear foci have been identified in candidate approaches (Lisby et al 2001Melo et al 2001;Zhu et al 2008;Germann et al 2011) and in genome-scale screens (Tkach et al 2012;Denervaud et al 2013;Mazumder et al 2013;Yu et al 2013). Nuclear foci are commonly thought of as centers of DNA repair, in part because foci formed by recombination repair proteins colocalize with double-strand DNA breaks (Lisby et al 2003).…”

Section: Resultsmentioning

confidence: 99%

“…The DUF2439 domain is also found in Dbl2 (Yu et al 2013), but does not appear to be important for DNA damage resistance or for nuclear focus formation. Further, ZGRF1 is likely membrane anchored and so might not be a true ortholog of Mte1.…”

Section: Orthologs Of Mte1mentioning

confidence: 99%

“…MTE1 appears to be an ortholog of the Schizosaccharomyces pombe dbl2 + gene (Yu et al 2013). Dbl2 colocalizes with the fission yeast Rad52, and with double-strand breaks, and is important for nuclear focus formation by Fml1, the fission yeast ortholog of Mph1 (Yu et al 2013).…”

Section: Orthologs Of Mte1mentioning

confidence: 99%

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MTE1 Functions with MPH1 in Double-Strand Break Repair

et al. 2016

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Double-strand DNA breaks occur upon exposure of cells to ionizing radiation and certain chemical agents or indirectly through replication fork collapse at DNA damage sites. If left unrepaired, double-strand breaks can cause genome instability and cell death, and their repair can result in loss of heterozygosity. In response to DNA damage, proteins involved in double-strand break repair by homologous recombination relocalize into discrete nuclear foci. We identified 29 proteins that colocalize with recombination repair protein Rad52 in response to DNA damage. Of particular interest, Ygr042w/Mte1, a protein of unknown function, showed robust colocalization with Rad52. Mte1 foci fail to form when the DNA helicase gene MPH1 is absent. Mte1 and Mph1 form a complex and are recruited to double-strand breaks in vivo in a mutually dependent manner. MTE1 is important for resolution of Rad52 foci during double-strand break repair and for suppressing break-induced replication. Together our data indicate that Mte1 functions with Mph1 in double-strand break repair.KEYWORDS DNA repair; recombination; double-strand breaks; break-induced replication; loss of heterozygosity; nuclear foci E FFECTIVE repair of double-strand DNA breaks (DSBs) is critical to the preservation of genome stability, yet most modes of DSB repair have significant potential to generate sequence alterations or sequence loss. Repair of DSBs by homologous recombination can result in loss of heterozygosity when resolution of recombination intermediates between homologous chromosomes results in a crossover. As such, cells possess several mechanisms by which crossing over can be suppressed in favor of noncrossover recombination products. Double Holliday junction (dHJ) intermediates that result from invasion of a homologous chromosome by both ends of a resected DSB (Szostak et al. 1983) can be resolved nucleolytically by the action of the Yen1 and Mus81/Mms4 endonucleases (Blanco et al. 2010;Ho et al. 2010) to produce a random distribution of crossover and noncrossover products. By contrast, the same dHJ intermediates can be dissolved by the combined helicase and ssDNA decatenase action of the Bloom/TopIIIa/Rmi1 complex (Sgs1/Top3/Rmi1 in yeast) (Wu et al. 2006;Yang et al. 2010) to yield exclusively noncrossover products (Wu and Hickson 2003). Crossovers can also be prevented if the D-loop structure that results from the first strand invasion by one end of a resected DSB into the homologous chromosome is unwound before capture of the second end to form the dHJ. Unwinding of D-loops is catalyzed in vitro and in vivo by the 39-to-59 DNA helicase Mph1 (Sun et al. 2008;Prakash et al. 2009) to prevent loss of heterozygosity due to crossovers and break-induced replication (BIR) (Luke-Glaser and Luke 2012;Mazon and Symington 2013;Stafa et al. 2014).The Mph1 DNA helicase was first identified as a deletion mutant with an increased mutation frequency (Entian et al. 1999). Subsequent characterization revealed that mph1 mutants are sensitive to the alkylating agent M...

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Section: Resultssupporting

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: Orthologs Of Mte1mentioning

confidence: 99%

Section: Orthologs Of Mte1mentioning

confidence: 99%

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MTE1 Functions with MPH1 in Double-Strand Break Repair

et al. 2016

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“…Strains expressing ER-GFP (called GFP-AHDL in previous publications) were provided by Snezhana Oliferenko (King's College London, London, UK) (Zhang et al, 2010). Strains expressing PX-BAR proteins fused with the YFP-FLAG-His6 (YFH) tag from their native promoters were constructed by using an overlap-extension PCR approach (Yu et al, 2013). Strains expressing CFP-Atg8 and Tdh1-YFH were as reported previously (Sun et al, 2013).…”

Section: Fission Yeast Strains and Mediamentioning

confidence: 99%

Atg20- and Atg24-family proteins promote organelle autophagy in fission yeast

Zhao

Liu

et al. 2016

Journal of Cell Science

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Autophagy cargos include not only soluble cytosolic materials but also bulky organelles, such as ER and mitochondria. In budding yeast, two proteins that contain the PX domain and the BAR domain, Atg20 and Atg24 (also known as Snx42 and Snx4, respectively) are required for organelle autophagy and contribute to general autophagy in a way that can be masked by compensatory mechanisms. It remains unclear why these proteins are important for organelle autophagy. Here, we show that in a distantly related fungal organism, the fission yeast Schizosaccharomyces pombe, autophagy of ER and mitochondria is induced by nitrogen starvation and is promoted by three Atg20-and Atg24-family proteins -Atg20, Atg24 and SPBC1711.11 (named here as Atg24b). These proteins localize at the pre-autophagosomal structure, or phagophore assembly site (PAS), during starvation. S. pombe Atg24 forms a homo-oligomer and acts redundantly with Atg20 and Atg24b, and the latter two proteins can form a hetero-oligomer. The organelle autophagy defect caused by the loss of these proteins is associated with a reduction of autophagosome size and a decrease in Atg8 accumulation at the PAS. These results provide new insights into the autophagic function of Atg20-and Atg24-family proteins.

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Modulation of meiotic homologous recombination by DNA helicases

Lorenz

2017

Yeast

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DNA helicases are ATP-driven motor proteins which translocate along DNA capable of dismantling DNA-DNA interactions and/or removing proteins bound to DNA. These biochemical capabilities make DNA helicases main regulators of crucial DNA metabolic processes, including DNA replication, DNA repair, and genetic recombination. This budding topic will focus on reviewing the function of DNA helicases important for homologous recombination during meiosis, and discuss recent advances in how these modulators of meiotic recombination are themselves regulated. The emphasis is placed on work in the two model yeasts, Saccharomyces cerevisiae and Schizosaccharomyces pombe, which has vastly expanded our understanding of meiotic homologous recombination, a process whose correct execution is instrumental for healthy gamete formation, and thus functioning sexual reproduction. Copyright © 2016 John Wiley & Sons, Ltd.

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A proteome-wide visual screen identifies fission yeast proteins localizing to DNA double-strand breaks

Cited by 37 publications

References 67 publications

MTE1 Functions with MPH1 in Double-Strand Break Repair

MTE1 Functions with MPH1 in Double-Strand Break Repair

Atg20- and Atg24-family proteins promote organelle autophagy in fission yeast

Modulation of meiotic homologous recombination by DNA helicases

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