Template Switching During Break-Induced Replication Is Promoted by the Mph1 Helicase in Saccharomyces cerevisiae

Chromosomal double-strand breaks (DSBs) that have only one end with homology to a donor duplex undergo repair by strand invasion followed by replication to the chromosome terminus (break-induced replication, BIR). Using a transformation-based assay system, it was previously shown that BIR could occur by several rounds of strand invasion, DNA synthesis, and dissociation. Here we describe a modification of the transformation-based assay to facilitate detection of switching between donor templates during BIR by genetic selection in diploid yeast. In addition to the expected recovery of template switch products, we found a high frequency of recombination between chromosome homologs during BIR, suggesting transfer of the DSB from the transforming linear DNA to the donor chromosome, initiating secondary recombination events. The frequency of BIR increased in the mph1Δ mutant, but the percentage of template switch events was significantly decreased, revealing an important role for Mph1 in promoting BIR-associated template switching. In addition, we show that the Mus81, Rad1, and Yen1 structure-selective nucleases act redundantly to facilitate BIR.

• Publication year

  2014

• Venue

  Genetics

• Publication date

  2014-02-04

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1534/genetics.114.162297PMID 24496010PMCID 3982708
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Migrating bubble during break-induced replication drives conservative DNA synthesis

  2013influential reference
• Heteroduplex DNA Position Defines the Roles of the Sgs1, Srs2, and Mph1 Helicases in Promoting Distinct Recombination Outcomes

  2013cited by this paper
• Mph1 and Mus81-Mms4 prevent aberrant processing of mitotic recombination intermediates.

  2013cited by this paper
• Break-induced replication occurs by conservative DNA synthesis

  2013cited by this paper
• Pif1 helicase and Polδ promote recombination-coupled DNA synthesis via bubble migration

  2013influential reference
• Pif1 family helicases suppress genome instability at G-quadruplex motifs

  2013cited by this paper
• Telomerase-Null Survivor Screening Identifies Novel Telomere Recombination Regulators

  2013influential reference
• Distinct Roles of Mus81, Yen1, Slx1-Slx4, and Rad1 Nucleases in the Repair of Replication-Born Double-Strand Breaks by Sister Chromatid Exchange

  2012cited by this paper
• The Mph1 Helicase Can Promote Telomere Uncapping and Premature Senescence in Budding Yeast

  2012cited by this paper
• Complex Chromosomal Rearrangements Mediated by Break-Induced Replication Involve Structure-Selective Endonucleases

  2012cited by this paper
• The Rad1–Rad10 nuclease promotes chromosome translocations between dispersed repeats

  2012cited by this paper
• Erratum to: Processing of joint molecule intermediates by structure-selective endonucleases during homologous recombination in eukaryotes

  2011cited by this paper
• Break-Induced Replication Is Highly Inaccurate

  2011cited by this paper
• The role of Holliday junction resolvases in the repair of spontaneous and induced DNA damage

  2011cited by this paper
• Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities

  2011cited by this paper
• Processing of joint molecule intermediates by structure-selective endonucleases during homologous recombination in eukaryotes

  2011cited by this paper
• Extensive DNA End Processing by Exo1 and Sgs1 Inhibits Break-Induced Replication

  2010cited by this paper
• Functional overlap between the structure-specific nucleases Yen1 and Mus81-Mms4 for DNA-damage repair in S. cerevisiae.

  2010cited by this paper
• Defective Resection at DNA Double-Strand Breaks Leads to De Novo Telomere Formation and Enhances Gene Targeting

  2010cited by this paper
• Mechanism of Holliday junction resolution by the human GEN1 protein.

  2010cited by this paper
• Mph1 requires mismatch repair-independent and -dependent functions of MutSα to regulate crossover formation during homologous recombination repair

  2010cited by this paper
• Mus81 and Yen1 promote reciprocal exchange during mitotic recombination to maintain genome integrity in budding yeast.

  2010cited by this paper
• Unwinding the functions of the Pif1 family helicases.

  2010cited by this paper
• Aberrant Double-Strand Break Repair Resulting in Half Crossovers in Mutants Defective for Rad51 or the DNA Polymerase δ Complex

  2009cited by this paper
• A tale of tails: insights into the coordination of 3′ end processing during homologous recombination

  2009cited by this paper
• Chromosomal Translocations Caused by Either Pol32-Dependent or Pol32-Independent Triparental Break-Induced Replication

  2009cited by this paper
• A Microhomology-Mediated Break-Induced Replication Model for the Origin of Human Copy Number Variation

  2009cited by this paper
• The Yeast Pif1 Helicase Prevents Genomic Instability Caused by G-Quadruplex-Forming CEB1 Sequences In Vivo

  2009cited by this paper
• Yeast Mph1 helicase dissociates Rad51-made D-loops: implications for crossover control in mitotic recombination.

  2009cited by this paper
• Defective Break-Induced Replication Leads to Half-Crossovers in Saccharomyces cerevisiae

  2008cited by this paper
• Break-induced replication: What is it and what is it for?

  2008cited by this paper
• The FANCM Ortholog Fml1 Promotes Recombination at Stalled Replication Forks and Limits Crossing Over during DNA Double-Strand Break Repair

  2008cited by this paper
• Role of the Saccharomyces cerevisiae Rad51 Paralogs in Sister Chromatid Recombination

  2008cited by this paper
• Template switching during break-induced replication

  2007cited by this paper
• Break-induced replication and telomerase-independent telomere maintenance require Pol32

  2007cited by this paper
• The Absence of Top3 Reveals an Interaction Between the Sgs1 and Pif1 DNA Helicases in Saccharomyces cerevisiae

  2006cited by this paper
• Break-induced replication and recombinational telomere elongation in yeast.

  2006cited by this paper
• Selection and analysis of spontaneous reciprocal mitotic cross-overs in Saccharomyces cerevisiae

  2006cited by this paper
• Conservative Inheritance of Newly Synthesized DNA in Double-Strand Break-Induced Gene Conversion

  2006cited by this paper
• The yeast Pif1p helicase removes telomerase from telomeric DNA

  2005cited by this paper
• RAD51-Dependent Break-Induced Replication Differs in Kinetics and Checkpoint Responses from RAD51-Mediated Gene Conversion

  2005cited by this paper
• RAD51-Dependent Break-Induced Replication in Yeast

  2004cited by this paper
• Slx1-Slx4 is a second structure-specific endonuclease functionally redundant with Sgs1-Top3.

  2003cited by this paper
• Generating crossovers by resolution of nicked Holliday junctions: a role for Mus81-Eme1 in meiosis.

  2003cited by this paper
• The Bloom's syndrome helicase suppresses crossing over during homologous recombination

  2003cited by this paper
• Srs2 and Sgs1-Top3 suppress crossovers during double-strand break repair in yeast.

  2003cited by this paper
• An Age-Induced Switch to a Hyper-Recombinational State

  2003cited by this paper
• Role of RAD 52 Epistasis Group Genes in Homologous Recombination and Double-Strand Break Repair

  2002cited by this paper
• Role of RAD52 Epistasis Group Genes in Homologous Recombination and Double-Strand Break Repair

  2002cited by this paper
• A molecular genetic dissection of the evolutionarily conserved N terminus of yeast Rad52.

  2002influential reference
• Fate of mat1 DNA strands during mating‐type switching in fission yeast

  2000cited by this paper
• DNA mismatch repair and genetic instability.

  2000cited by this paper
• Multiple heterologies increase mitotic double-strand break-induced allelic gene conversion tract lengths in yeast.

  1999cited by this paper
• Chromosome break-induced DNA replication leads to nonreciprocal translocations and telomere capture.

  1998cited by this paper
• "Break copy" duplication: a model for chromosome fragment formation in Saccharomyces cerevisiae.

  1997cited by this paper
• Recombinational repair of gaps in DNA is asymmetric in Ustilago maydis and can be explained by a migrating D-loop model.

  1996cited by this paper
• Plasmid-mediated induction of recombination in yeast.

  1994influential reference
• The saccharomyces PIF1 DNA helicase inhibits telomere elongation and de novo telomere formation.

  1994influential reference
• Efficient copying of nonhomologous sequences from ectopic sites via P-element-induced gap repair

  1994cited by this paper
• Removal of nonhomologous DNA ends in double-strand break recombination: the role of the yeast ultraviolet repair gene RAD1.

  1992cited by this paper
• A DNA double chain break stimulates triparental recombination in Saccharomyces cerevisiae.

  1989cited by this paper
• Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis.

  1984cited by this paper
• A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance

  1984cited by this paper
• Transformation of intact yeast cells treated with alkali cations.

  1984cited by this paper
• The double-strand-break repair model for recombination.

  1983cited by this paper
• Methods in yeast genetics

  1979cited by this paper

• Elevated mutation in haploid yeast driven by translesion synthesis

  2026cites this paper
• Mechanisms and genomic implications of break-induced replication

  2025cites this paper
• Atypical R-loops in cancer: decoding molecular chaos for therapeutic gain

  2025cites this paper
• Late steps of allelic break-induced replication suppress tandem duplication associated with BRCA1 deficiency

  2025cites this paper
• Stressed? Break-induced replication comes to the rescue!

  2024cites this paper
• Induction of homologous recombination by site-specific replication stress.

  2024cites this paper
• FANCM branchpoint translocase: Master of traverse, reverse and adverse DNA repair.

  2024cites this paper
• Break-induced replication orchestrates resection-dependent template switching

  2023cites this paper
• Delineation of two multi-invasion-induced rearrangement pathways that differently affect genome stability

  2023cites this paper
• Delineation of two multi-invasion-induced rearrangement pathways that differently affect genome stability

  2023influential citation
• RTEL-1 and DNA polymerase theta promote subtelomeric DNA synthesis and telomere fusion in C. elegans

  2022cites this paper
• Mechanism for inverted-repeat recombination induced by a replication fork barrier

  2022cites this paper
• Measuring the contributions of helicases to break-induced replication.

  2022cites this paper
• Break-induced replication: unraveling each step.

  2022cites this paper
• Mechanism for inverted-repeat recombination induced by a replication fork barrier

  2022cites this paper
• Repair of DNA Breaks by Break-Induced Replication.

  2021cites this paper
• Take a Break to Repair: A Dip in the World of Double-Strand Break Repair Mechanisms Pointing the Gaze on Archaea

  2021cites this paper
• Homologous recombination induced by a replication fork barrier requires cooperation between strand invasion and strand annealing activities

  2021cites this paper
• The dark side of homology-directed repair.

  2021cites this paper
• DNA Damage Tolerance in the Yeast Saccharomyces cerevisiae

  2021cites this paper
• Stimulation of adaptive gene amplification by origin firing under replication fork constraint

  2021cites this paper
• Signaling pathways involved in cell cycle arrest during the DNA breaks.

  2021cites this paper
• Mechanisms restraining break‐induced replication at two‐ended DNA double‐strand breaks

  2021cites this paper
• Double-strand breaks induce short-scale DNA replication and damage amplification in the fully grown mouse oocytes

  2021cites this paper
• Interstitial telomere sequences disrupt break-induced replication and drive formation of ectopic telomeres

  2020cites this paper
• Saccharomyces cerevisiae Mus81-Mms4 prevents accelerated senescence in telomerase-deficient cells

  2020cites this paper
• DNA Helicase Mph1FANCM Ensures Meiotic Recombination between Parental Chromosomes by Dissociating Precocious Displacement Loops.

  2020cites this paper
• Mechanisms preventing Break-Induced Replication during repair of two-ended DNA double-strand breaks

  2020cites this paper
• FANCM regulates repair pathway choice at stalled replication forks

  2020cites this paper
• Leveraging DNA Damage Response Pathways to Enhance the Precision of CRISPR-Mediated Genome Editing

  2020cites this paper
• DNA Polymerase Delta Synthesizes Both Strands during Break-Induced Replication.

  2019cites this paper
• Homologous Recombination and the Formation of Complex Genomic Rearrangements.

  2019cites this paper
• Factors affecting template switch recombination associated with restarted DNA replication

  2019influential citation
• At the Beginning of the End and in the Middle of the Beginning: Structure and Maintenance of Telomeric DNA Repeats and Interstitial Telomeric Sequences

  2019cites this paper
• Moving forward one step back at a time: reversibility during homologous recombination

  2019cites this paper
• DNA double-strand break repair-pathway choice in somatic mammalian cells

  2019cites this paper
• Interstitial Telomere Sequences Disrupt Break Induced Replication

  2019influential citation
• The biochemistry of early meiotic recombination intermediates

  2018cites this paper
• Loss of Cohesin Subunit Rec8 Switches Rad51 Mediator Dependence in Resistance to Formaldehyde Toxicity in Ustilago maydis

  2018cites this paper
• Dynamic Processing of Displacement Loops During Recombinational DNA Repair

  2018influential citation
• Multi‐Invasion‐Induced Rearrangements as a Pathway for Physiological and Pathological Recombination

  2018cites this paper
• Break‐induced replication links microsatellite expansion to complex genome rearrangements

  2017cites this paper
• Role of PCNA and RFC in promoting Mus81-complex activity

  2017cites this paper
• Homology search guidance by the yeast recombination enhancer

  2017cites this paper
• In Saccharomyces cerevisiae gene targeting fidelity depends on a transformation method and proportion of the overall length of the transforming and targeted DNA

  2017cites this paper
• Break induced replication in eukaryotes: mechanisms, functions, and consequences

  2017cites this paper
• Multi-invasions Are Recombination Byproducts that Induce Chromosomal Rearrangements.

  2017influential citation
• Not Restricted to the Ends: Yeast Telomere Proteins Rif1 and Cdc13 Function in Double-Strand Break Repair Pathways with Implications for Genome Stability

  2016cites this paper
• Assembly of Slx4 Checkpoint Signaling Complexes Behind Stressed DNA Replication Forks

  2016cites this paper
• DISSERTATION INVESTIGATION OF MECHANISMS OF MITOTIC RECOMBINATION IN YEAST Submitted by Lisa Victoria Harcy Graduate Degree Program in Cell and Molecular Biology In partial fulfillment of the requirements For the Degree of Doctor of Philosophy

  2016cites this paper
• MTE1 Functions with MPH1 in Double-Strand Break Repair

  2016influential citation
• RPA Stabilization of Single-Stranded DNA Is Critical for Break-Induced Replication.

  2016cites this paper
• Repair Pathway Choices and Consequences at the Double-Strand Break.

  2016cites this paper
• Error‐Prone Repair of DNA Double‐Strand Breaks

  2016cites this paper
• Pfh1 Is an Accessory Replicative Helicase that Interacts with the Replisome to Facilitate Fork Progression and Preserve Genome Integrity

  2016cites this paper
• Sgs1 and Mph1 Helicases Enforce the Recombination Execution Checkpoint During DNA Double-Strand Break Repair in Saccharomyces cerevisiae

  2016cites this paper
• Biochemical Activities and Genetic Functions of the Drosophila melanogaster Fancm Helicase in DNA Repair

  2016cites this paper
• Choices have consequences: the nexus between DNA repair pathways and genomic instability in cancer

  2016cites this paper
• Eukaryotic DNA Polymerases in Homologous Recombination.

  2016cites this paper
• Functions and regulation of the multitasking FANCM family of DNA motor proteins

  2015cites this paper
• YGR042W/MTE1 Functions in Double-Strand Break Repair with MPH1

  2015cites this paper
• Translesion Polymerases Drive Microhomology-Mediated Break-Induced Replication Leading to Complex Chromosomal Rearrangements.

  2015cites this paper
• Recombination occurs within minutes of replication blockage by RTS1 producing restarted forks that are prone to collapse

  2015influential citation
• An investigation into the initiation of VSG switching in Trypanosoma brucei

  2015cites this paper
• Perinuclear tethers license telomeric DSBs for a broad kinesin- and NPC-dependent DNA repair process

  2015cites this paper
• Chromosome rearrangements via template switching between diverged repeated sequences

  2014cites this paper
• Understanding the mechanisms underlying DSB repair-induced mutagenesis at distant loci in yeast

  2014cites this paper
• Mechanisms and Regulation of Mitotic Recombination in Saccharomyces cerevisiae

  2014cites this paper
• Frequent Interchromosomal Template Switches during Gene Conversion in S. cerevisiae.

  2014cites this paper
• Sgs1 and Exo1 suppress targeted chromosome duplication during ends-in and ends-out gene targeting.

  2014cites this paper