Fine-Structure Mapping of Meiosis-Specific Double-Strand DNA Breaks at a Recombination Hotspot Associated With an Insertion of Telomeric Sequences Upstream of the HIS4 Locus in Yeast

Abstract Meiotic recombination in Saccharomyces cerevisiae is initiated by double-strand DNA breaks (DSBs). Using two approaches, we mapped the position of DSBs associated with a recombination hotspot created by insertion of telomeric sequences into the region upstream of HIS4. We found that the breaks have no obvious sequence specificity and localize to a region of ~50 bp adjacent to the telomeric insertion. By mapping the breaks and by studies of the exonuclease III sensitivity of the broken ends, we conclude that most of the broken DNA molecules have blunt ends with 3′-hydroxyl groups.

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Mer2 binds directly to both nucleosomes and axial proteins as the keystone of meiotic recombination

10.1101/2020.07.30.228908 ◽

2020 ◽

Author(s):

Dorota Rousova ◽

Saskia K. Funk ◽

Heidi Reichle ◽

John R. Weir

Keyword(s):

Sexual Reproduction ◽

Meiotic Recombination ◽

Dna Breaks ◽

Homologous Chromosomes ◽

Protein Biochemistry ◽

Double Strand Dna Breaks ◽

Domain Protein ◽

Meiosis I ◽

Double Strand Dna

One of the defining features of sexual reproduction is the recombination events that take place during meiosis I. Recombination is both evolutionarily advantageous, but also mechanistically necessary to form the crossovers that link homologous chromosomes. Meiotic recombination is initiated through the placement of programmed double-strand DNA breaks (DSBs) mediated by the protein Spo11. The timing, number, and physical placement of DSBs are carefully controlled through a variety of protein machinery. Previous work has implicated Mer2(IHO1 in mammals) to be involved in both the placement of breaks, and their timing. In this study we use a combination of protein biochemistry and biophysics to extensively characterise various roles of the Mer2. We gain further insights into the details of Mer2 interaction with the PHD protein Spp1, reveal that Mer2 is a novel nucleosome binder, and suggest how Mer2’s interaction with the HORMA domain protein Hop1 (HORMAD1/2 in mammals) is controlled.

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Use of a ring chromosome and pulsed-field gels to study interhomolog recombination, double-strand DNA breaks and sister-chromatid exchange in yeast.

Genetics ◽

10.1093/genetics/123.4.695 ◽

1989 ◽

Vol 123 (4) ◽

pp. 695-713 ◽

Cited By ~ 4

Author(s):

J C Game ◽

K C Sitney ◽

V E Cook ◽

R K Mortimer

Keyword(s):

Sister Chromatid ◽

Meiotic Recombination ◽

Sister Chromatid Exchange ◽

Recombinant Dna ◽

Ring Chromosome ◽

X Rays ◽

Dna Breaks ◽

Pulsed Field ◽

Double Strand Dna Breaks ◽

Double Strand Dna

Abstract We describe a system that uses pulsed-field gels for the physical detection of recombinant DNA molecules, double-strand DNA breaks (DSB) and sister-chromatid exchange in the yeast Saccharomyces cerevisiae. The system makes use of a circular variant of chromosome III (Chr. III). Meiotic recombination between this ring chromosome and a linear homolog produces new molecules of sizes distinguishable on gels from either parental molecule. We demonstrate that these recombinant molecules are not present either in strains with two linear Chr. III molecules or in rad50 mutants, which are defective in meiotic recombination. In conjunction with the molecular endpoints, we present data on the timing of commitment to meiotic recombination scored genetically. We have used x-rays to linearize circular Chr. III, both to develop a sensitive method for measuring frequency of DSB and as a means of detecting double-sized circles originating in part from sister-chromatid exchange, which we find to be frequent during meiosis.

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Initiation of meiotic recombination by double-strand DNA breaks in S. pombe

Cell ◽

10.1016/0092-8674(86)90348-x ◽

1986 ◽

Vol 46 (5) ◽

pp. 725-731 ◽

Cited By ~ 64

Author(s):

Amar J.S. Klar ◽

Lisa M. Miglio

Keyword(s):

Meiotic Recombination ◽

Dna Breaks ◽

Double Strand Dna Breaks ◽

Double Strand Dna

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Competition Between Adjacent Meiotic Recombination Hotspots in the Yeast Saccharomyces cerevisiae

Genetics ◽

10.1093/genetics/145.3.661 ◽

1997 ◽

Vol 145 (3) ◽

pp. 661-670 ◽

Cited By ~ 1

Author(s):

Qing-Qing Fan ◽

Fei Xu ◽

Michael A White ◽

Thomas D Petes

Keyword(s):

Saccharomyces Cerevisiae ◽

Meiotic Recombination ◽

Telomeric Sequence ◽

Coding Region ◽

Dna Breaks ◽

Local Formation ◽

Yeast Saccharomyces Cerevisiae ◽

Recombination Hotspots ◽

Double Strand Dna Breaks ◽

His4 Gene

In a wild-type strain of Saccharomyces cerevisiae, a hotspot for meiotic recombination is located upstream of the HIS4 gene. An insertion of a 49-bp telomeric sequence into the coding region of HIS4 strongly stimulates meiotic recombination and the local formation of meiosis-specific double-strand DNA breaks (DSBs). When strains are constructed in which both hotspots are heterozygous, hotspot activity is substantially less when the hotspots are on the same chromosome than when they are on opposite chromosomes.

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Expression of double strand DNA breaks repair genes in pterygium

Ophthalmic Genetics ◽

10.3109/13816810.2010.524907 ◽

2010 ◽

Vol 32 (1) ◽

pp. 39-47 ◽

Cited By ~ 4

Author(s):

Anna Łękawa–Ilczuk ◽

Halina Antosz ◽

Beata Rymgayłło–Jankowska ◽

Tomasz Żarnowski

Keyword(s):

Dna Breaks ◽

Double Strand Dna Breaks ◽

Repair Genes ◽

Double Strand Dna

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Inside Back Cover: Double-Strand DNA Breaks Induced by Paracyclophane Gold(I) Complexes (Chem. Eur. J. 26/2017)

Chemistry - A European Journal ◽

10.1002/chem.201700517 ◽

2017 ◽

Vol 23 (26) ◽

pp. 6459-6459

Author(s):

Sebastian Bestgen ◽

Carmen Seidl ◽

Thomas Wiesner ◽

Andreas Zimmer ◽

Martina Falk ◽

...

Keyword(s):

Dna Breaks ◽

Back Cover ◽

Double Strand Dna Breaks ◽

Double Strand Dna

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Nucleosomes Suppress the Formation of Double-strand DNA Breaks during Attempted Base Excision Repair of Clustered Oxidative Damages

Journal of Biological Chemistry ◽

10.1074/jbc.m114.571588 ◽

2014 ◽

Vol 289 (29) ◽

pp. 19881-19893 ◽

Cited By ~ 28

Author(s):

Wendy J. Cannan ◽

Betty P. Tsang ◽

Susan S. Wallace ◽

David S. Pederson

Keyword(s):

Base Excision Repair ◽

Excision Repair ◽

Dna Breaks ◽

Oxidative Damages ◽

Double Strand Dna Breaks ◽

Base Excision ◽

Double Strand Dna

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Roles of Caenorhabditis elegans WRN Helicase in DNA Damage Responses, and a Comparison with Its Mammalian Homolog: A Mini-Review

Gerontology ◽

10.1159/000439200 ◽

2015 ◽

Vol 62 (3) ◽

pp. 296-303 ◽

Cited By ~ 6

Author(s):

Jin-Sun Ryu ◽

Hyeon-Sook Koo

Keyword(s):

Dna Damage ◽

Caenorhabditis Elegans ◽

Werner Syndrome ◽

Model Organisms ◽

Current Status ◽

Helicase Domain ◽

Replication Forks ◽

Dna Breaks ◽

Double Strand Dna Breaks ◽

Double Strand Dna

Werner syndrome protein (WRN) is unusual among RecQ family DNA helicases in having an additional exonuclease activity. WRN is involved in the repair of double-strand DNA breaks via the homologous recombination and nonhomologous end joining pathways, and also in the base excision repair pathway. In addition, the protein promotes the recovery of stalled replication forks. The helicase activity is thought to unwind DNA duplexes, thereby moving replication forks or Holliday junctions. The targets of the exonuclease could be the nascent DNA strands at a replication fork or the ends of double-strand DNA breaks. However, it is not clear which enzyme activities are essential for repairing different types of DNA damage. Model organisms such as mice, flies, and worms deficient in WRN homologs have been investigated to understand the physiological results of defects in WRN activity. Premature aging, the most remarkable characteristic of Werner syndrome, is also seen in the mutant mice and worms, and hypersensitivity to DNA damage has been observed in WRN mutants of all three model organisms, pointing to conservation of the functions of WRN. In the nematode Caenorhabditis elegans, the WRN homolog contains a helicase domain but no exonuclease domain, so that this animal is very useful for studying the in vivo functions of the helicase without interference from the activity of the exonuclease. Here, we review the current status of investigations of C. elegans WRN-1 and discuss its functional differences from the mammalian homologs.

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