scholarly journals Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae

1999 ◽  
Vol 63 (2) ◽  
pp. 349-404 ◽  
Author(s):  
Frédéric Pâques ◽  
James E. Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Repair ◽  
Genetic Recombination ◽  
Double Strand Breaks ◽  
Molecular Models ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks ◽  
The Past ◽  
Biochemical Studies

SUMMARY The budding yeast Saccharomyces cerevisiae has been the principal organism used in experiments to examine genetic recombination in eukaryotes. Studies over the past decade have shown that meiotic recombination and probably most mitotic recombination arise from the repair of double-strand breaks (DSBs). There are multiple pathways by which such DSBs can be repaired, including several homologous recombination pathways and still other nonhomologous mechanisms. Our understanding has also been greatly enriched by the characterization of many proteins involved in recombination and by insights that link aspects of DNA repair to chromosome replication. New molecular models of DSB-induced gene conversion are presented. This review encompasses these different aspects of DSB-induced recombination in Saccharomyces and attempts to relate genetic, molecular biological, and biochemical studies of the processes of DNA repair and recombination.

scholarly journals The essential DNA polymerases delta and epsilon are involved in repair of UV-damaged DNA in the yeast Saccharomyces cerevisiae.

1999 ◽  
Vol 46 (2) ◽  
pp. 289-298 ◽  
Author(s):  
A Hałas ◽  
Z Policińska ◽  
H Baranowska ◽  
W J Jachymczyk
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Repair ◽  
Uv Irradiation ◽  
Dna Polymerases ◽  
Relative Molecular Mass ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks ◽  
Single Strand Breaks ◽  
Mutant Strains ◽  
Cellular Dna

We have studied the ability of yeast DNA polymerases to carry out repair of lesions caused by UV irradiation in Saccharomyces cerevisiae. By the analysis of postirradiation relative molecular mass changes in cellular DNA of different DNA polymerases mutant strains, it was established that mutations in DNA polymerases delta and epsilon showed accumulation of single-strand breaks indicating defective repair. Mutations in other DNA polymerase genes exhibited no defects in DNA repair. Thus, the data obtained suggest that DNA polymerases delta and epsilon are both necessary for DNA replication and for repair of lesions caused by UV irradiation. The results are discussed in the light of current concepts concerning the specificity of DNA polymerases in DNA repair.

scholarly journals The Drosophila melanogaster Ortholog of RFWD3 Functions Independently of RAD51 During DNA Repair

2020 ◽  
Vol 10 (3) ◽  
pp. 999-1004
Author(s):  
Juan Carvajal-Garcia ◽  
Evan R. Gales ◽  
Dale A. Ramsden ◽  
Jeff Sekelsky
Keyword(s):  
Dna Repair ◽  
Drosophila Melanogaster ◽  
Ring Finger ◽  
Double Strand Breaks ◽  
Specific Gene ◽  
Genetic Screens ◽  
Strand Breaks ◽  
Ancestral Function ◽  
Human Ortholog

Repair of damaged DNA is required for the viability of all organisms. Studies in Drosophila melanogaster, driven by the power of genetic screens, pioneered the discovery and characterization of many genes and pathways involved in DNA repair in animals. However, fewer than half of the alleles identified in these screens have been mapped to a specific gene, leaving a potential for new discoveries in this field. Here we show that the previously uncharacterized mutagen sensitive gene mus302 codes for the Drosophila melanogaster ortholog of the E3 ubiquitin ligase RING finger and WD domain protein 3 (RFWD3). In human cells, RFWD3 promotes ubiquitylation of RPA and RAD51 to facilitate repair of collapsed replication forks and double-strand breaks through homologous recombination. Despite the high similarity in sequence to the human ortholog, our evidence fails to support a role for Mus302 in the repair of these types of damage. Last, we observe that the N-terminal third of RFWD3 is only found in mammals, but not in other vertebrates or invertebrates. We propose that the new N-terminal sequence accounts for the acquisition of a new biological function in mammals that explains the functional differences between the human and the fly orthologs, and that Drosophila Mus302 may retain the ancestral function of the protein.

Repair of double-strand breaks in plasmid DNA in the yeast Saccharomyces cerevisiae

1988 ◽  
Vol 213 (2-3) ◽  
pp. 421-424 ◽  
Author(s):  
Joseph R. Perera ◽  
Alexander V. Glasunov ◽  
Vadim M. Glaser ◽  
Alla V. Boreiko
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasmid Dna ◽  
Double Strand Breaks ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks

scholarly journals A General Method for Identifying Recessive Diploid-Specific Mutations in Saccharomyces cerevisiae, Its Application to the Isolation of Mutants Blocked at Intermediate Stages of Meiotic Prophase and Characterization of a New Gene SAE2

Genetics ◽  
1997 ◽  
Vol 146 (3) ◽  
pp. 797-816 ◽  
Author(s):  
Andrew H Z McKee ◽  
Nancy Kleckner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Prophase ◽  
Temperature Sensitive ◽  
Yeast Saccharomyces Cerevisiae ◽  
New Approach ◽  
Strand Breaks ◽  
Recessive Mutations ◽  
New Genes ◽  
New Gene

We describe a general new approach for identifying recessive mutations that affect diploid strains of yeast Saccharomyces cerevisiae and the application of this method to the identification of mutations that confer an intermediate block in meiotic prophase chromosome metabolism. The method uses a temperature-sensitive conjugation mutation ste7-1 in combination with homothallism. The mutations of interest confer a defect in spore formation that is dependent upon a gene required for initiation of meiotic recombination and development of meiosis-specific chromosome structure (SPO11). Identified in this screen were null mutations of the DMC1 gene, nonnull mutations of RAD50 (rad50S, and mutations in three new genes designated SAE1, SAE2 and SAE3 (Sporulation in the Absence of Spo Eleven). Molecular characterization of the SAE2 gene and characterization of meiotic and mitotic phenotypes of sae2 mutants are also presented. The phenotypes conferred by a sae2 null mutation are virtually indistinguishable from those conferred by the previously identified nonnull mutations of RAD50 (rad50S). Most notably, both mutations confer only weak sensitivity to the radiomimetic agent methyl methane sulfonate (MMS) but completely block resection and turnover of meiosis-specific double-strand breaks. These observations provide further evidence that this constellation of phenotypes identifies a specific molecular function.

scholarly journals Repair of Endonuclease-Induced Double-Strand Breaks in Saccharomyces cerevisiae: Essential Role for Genes Associated with Nonhomologous End-Joining

Genetics ◽  
1999 ◽  
Vol 152 (4) ◽  
pp. 1513-1529 ◽  
Author(s):  
L Kevin Lewis ◽  
James W Westmoreland ◽  
Michael A Resnick
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Excision Repair ◽  
Cell Killing ◽  
Double Strand Breaks ◽  
Nonhomologous End Joining ◽  
Chromosomal Dna ◽  
End Joining ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks

Abstract Repair of double-strand breaks (DSBs) in chromosomal DNA by nonhomologous end-joining (NHEJ) is not well characterized in the yeast Saccharomyces cerevisiae. Here we demonstrate that several genes associated with NHEJ perform essential functions in the repair of endonuclease-induced DSBs in vivo. Galactose-induced expression of EcoRI endonuclease in rad50, mre11, or xrs2 mutants, which are deficient in plasmid DSB end-joining and some forms of recombination, resulted in G2 arrest and rapid cell killing. Endonuclease synthesis also produced moderate cell killing in sir4 strains. In contrast, EcoRI caused prolonged cell-cycle arrest of recombination-defective rad51, rad52, rad54, rad55, and rad57 mutants, but cells remained viable. Cell-cycle progression was inhibited in excision repair-defective rad1 mutants, but not in rad2 cells, indicating a role for Rad1 processing of the DSB ends. Phenotypic responses of additional mutants, including exo1, srs2, rad5, and rdh54 strains, suggest roles in recombinational repair, but not in NHEJ. Interestingly, the rapid cell killing in haploid rad50 and mre11 strains was largely eliminated in diploids, suggesting that the cohesive-ended DSBs could be efficiently repaired by homologous recombination throughout the cell cycle in the diploid mutants. These results demonstrate essential but separable roles for NHEJ pathway genes in the repair of chromosomal DSBs that are structurally similar to those occurring during cellular development.

scholarly journals Hypermutability of Damaged Single-Strand DNA Formed at Double-Strand Breaks and Uncapped Telomeres in Yeast Saccharomyces cerevisiae

PLoS Genetics ◽  
2008 ◽  
Vol 4 (11) ◽  
pp. e1000264 ◽  
Author(s):  
Yong Yang ◽  
Joan Sterling ◽  
Francesca Storici ◽  
Michael A. Resnick ◽  
Dmitry A. Gordenin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Double Strand Breaks ◽  
Single Strand ◽  
Yeast Saccharomyces Cerevisiae ◽  
Strand Breaks

scholarly journals The Drosophila melanogaster ortholog of RFWD3 functions independently of RAD51 during DNA repair

2019 ◽  
Author(s):  
Juan Carvajal-Garcia ◽  
Evan R. Gales ◽  
Dale A. Ramsden ◽  
Jeff Sekelsky
Keyword(s):  
Dna Repair ◽  
Drosophila Melanogaster ◽  
Ring Finger ◽  
Double Strand Breaks ◽  
Specific Gene ◽  
Genetic Screens ◽  
Strand Breaks ◽  
Ancestral Function ◽  
Human Ortholog

AbstractRepair of damaged DNA is required for the viability of all organisms. Studies in Drosophila melanogaster, driven by the power of genetic screens, pioneered the discovery and characterization of many genes and pathways involved in DNA repair in animals. However, fewer than half of the alleles identified in these screens have been mapped to a specific gene, leaving a potential for new discoveries in this field. Here we show that the previously uncharacterized mutagen sensitive gene mus302 codes for the Drosophila melanogaster ortholog of the E3 ubiquitin ligase RING finger and WD domain protein 3 (RFWD3). In human cells, RFWD3 promotes ubiquitylation of RPA and RAD51 to facilitate repair of collapsed replication forks and double strand breaks through homologous recombination. Despite the high similarity in sequence to the human ortholog, our evidence fails to support a role for Mus302 in the repair of these types of damage. Last, we observe that the N-terminal third of RFWD3 is only present in mammals and absent in the rest of vertebrates and invertebrates. We propose that the additional N-terminal portion accounts for the acquisition of a new biological function in mammals that explains the functional differences between the human and the fly orthologs, and that Drosophila Mus302 may retain the ancestral function of the protein.

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