scholarly journals Genetic evidence that the meiotic recombination hotspot at the HIS4 locus of Saccharomyces cerevisiae does not represent a site for a symmetrically processed double-strand break.

Genetics ◽  
1993 ◽  
Vol 134 (1) ◽  
pp. 5-19 ◽  
Author(s):  
S E Porter ◽  
M A White ◽  
T D Petes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Site ◽  
Meiotic Recombination ◽  
Strand Break ◽  
Yeast Saccharomyces Cerevisiae ◽  
Recombination Hotspot ◽  
Dna Break ◽  
A Site ◽  
Aberrant Segregation ◽  
His4 Gene

Abstract In the yeast Saccharomyces cerevisiae, the binding of the Rap1 protein to a site located between the 5' end of the HIS4 gene and the 3' end of BIK1 stimulates meiotic recombination at both flanking loci. By using strains that contain mutations located in HIS4 and BIK1, we found that most recombination events stimulated by the binding of Rap1 involve HIS4 or BIK1, rather than bidirectional events including both loci. The patterns of aberrant segregation indicate that most of the Rap1-stimulated recombination events do not represent the symmetric processing of a double-strand DNA break located at the Rap1-binding site.

scholarly journals Crossover Interference in Saccharomyces cerevisiae Requires a TID1/RDH54- and DMC1-Dependent Pathway

Genetics ◽  
2003 ◽  
Vol 163 (4) ◽  
pp. 1273-1286 ◽  
Author(s):  
Miki Shinohara ◽  
Kazuko Sakai ◽  
Akira Shinohara ◽  
Douglas K Bishop
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
Strand Break ◽  
Tetrad Analysis ◽  
Yeast Saccharomyces Cerevisiae ◽  
Crossover Interference ◽  
Meiotic Progression ◽  
Invasion Stage ◽  
Strand Invasion ◽  
Dependent Pathway

Abstract Two RecA-like recombinases, Rad51 and Dmc1, function together during double-strand break (DSB)-mediated meiotic recombination to promote homologous strand invasion in the budding yeast Saccharomyces cerevisiae. Two partially redundant proteins, Rad54 and Tid1/Rdh54, act as recombinase accessory factors. Here, tetrad analysis shows that mutants lacking Tid1 form four-viable-spore tetrads with levels of interhomolog crossover (CO) and noncrossover recombination similar to, or slightly greater than, those in wild type. Importantly, tid1 mutants show a marked defect in crossover interference, a mechanism that distributes crossover events nonrandomly along chromosomes during meiosis. Previous work showed that dmc1Δ mutants are strongly defective in strand invasion and meiotic progression and that these defects can be partially suppressed by increasing the copy number of RAD54. Tetrad analysis is used to show that meiotic recombination in RAD54-suppressed dmc1Δ cells is similar to that in tid1; the frequency of COs and gene conversions is near normal, but crossover interference is defective. These results support the proposal that crossover interference acts at the strand invasion stage of recombination.

scholarly journals Competition Between Adjacent Meiotic Recombination Hotspots in the Yeast Saccharomyces cerevisiae

Genetics ◽  
1997 ◽  
Vol 145 (3) ◽  
pp. 661-670 ◽  
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.

scholarly journals Properties of Natural Double-Strand-Break Sites at a Recombination Hotspot in Saccharomyces cerevisiae

Genetics ◽  
2003 ◽  
Vol 165 (1) ◽  
pp. 101-114 ◽  
Author(s):  
Stuart J Haring ◽  
George R Halley ◽  
Alex J Jones ◽  
Robert E Malone
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Meiotic Recombination ◽  
Strand Break ◽  
Double Strand Break ◽  
Recombination Hotspot ◽  
Recombination Hotspots ◽  
Naturally Occurring ◽  
High Level ◽  
Site Recognition

Abstract This study addresses three questions about the properties of recombination hotspots in Saccharomyces cerevisiae: How much DNA is required for double-strand-break (DSB) site recognition? Do naturally occurring DSB sites compete with each other in meiotic recombination? What role does the sequence located at the sites of DSBs play? In S. cerevisiae, the HIS2 meiotic recombination hotspot displays a high level of gene conversion, a 3′-to-5′ conversion gradient, and two DSB sites located ∼550 bp apart. Previous studies of hotspots, including HIS2, suggest that global chromosome structure plays a significant role in recombination activity, raising the question of how much DNA is sufficient for hotspot activity. We find that 11.5 kbp of the HIS2 region is sufficient to partially restore gene conversion and both DSBs when moved to another yeast chromosome. Using a variety of different constructs, studies of hotspots have indicated that DSB sites compete with one another for DSB formation. The two naturally occurring DSBs at HIS2 afforded us the opportunity to examine whether or not competition occurs between these native DSB sites. Small deletions of DNA at each DSB site affect only that site; analyses of these deletions show no competition occurring in cis or in trans, indicating that DSB formation at each site at HIS2 is independent. These small deletions significantly affect the frequency of DSB formation at the sites, indicating that the DNA sequence located at a DSB site can play an important role in recombination initiation.

Analysis of a gene conversion gradient at the HIS4 locus in Saccharomyces cerevisiae.

Genetics ◽  
1992 ◽  
Vol 132 (1) ◽  
pp. 113-123 ◽  
Author(s):  
P Detloff ◽  
M A White ◽  
T D Petes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Mismatch Repair ◽  
Meiotic Recombination ◽  
Present Evidence ◽  
Homologous Chromosomes ◽  
A Site ◽  
His4 Gene ◽  
Heteroduplex Formation ◽  
Recombination Gene

Abstract Heteroduplexes formed between genes on homologous chromosomes are intermediates in meiotic recombination. In the HIS4 gene of Saccharomyces cerevisiae, most mutant alleles at the 5' end of the gene have a higher rate of meiotic recombination (gene conversion) than mutant alleles at the 3' end of the gene. Such gradients are usually interpreted as indicating a higher frequency of heteroduplex formation at the high conversion end of the gene. We present evidence indicating that the gradient of conversion at HIS4 primarily reflects the direction of mismatch repair rather than the frequency of heteroduplex formation. We also identify a site located between the 5' end of HIS4 and the 3' end of BIK1 that stimulates heteroduplex formation at HIS4 and BIK1.

Analysis of Meiotic Recombination Pathways in the Yeast Saccharomyces cerevisiae

Genetics ◽  
1996 ◽  
Vol 144 (1) ◽  
pp. 71-86 ◽  
Author(s):  
Yang Mao-Draayer ◽  
Anne M Galbraith ◽  
Douglas L Pittman ◽  
Marc Cool ◽  
Robert E Malone
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
The Other ◽  
Yeast Saccharomyces Cerevisiae ◽  
Recombination Hotspot ◽  
Early Genes ◽  
Intrachromosomal Recombination ◽  
Mutant Phenotypes ◽  
Recombination Pathway

Abstract In the yeast, Saccharomyces cerevisiae, several genes appear to act early in meiotic recombination. HOPl and REDl have been classified as such early genes. The data in this paper demonstrate that neither a redl nor a hopl mutation can rescue the inviable spores produced by a rad52 spol3 strain; this phenotype helps to distinguish these two genes from other early meiotic recombination genes such as SPOll, REC104, or A4EI4. In contrast, either a redl or a hopl mutation can rescue a rad50S spol3 strain; this phenotype is similar to that conferred by mutations in the other early recombination genes (e.g., REC104). These two different results can be explained because the data presented here indicate that a rad50S mutation does not diminish meiotic intrachromosomal recombination, similar to the mutant phenotypes conferred by redl or hopl. Of course, REDl and HOPl do act in the normal meiotic interchromosomal recombination pathway; they reduce interchromosomal recombination to ~10% of normal levels. We demonstrate that a mutation in a gene (REC104) required for initiation of exchange is completely epistatic to a mutation in REDl. Finally, mutations in either HOPl or RED1 reduce the number of doublestrand breaks observed at the HIS2 meiotic recombination hotspot.

scholarly journals Measurements of excision repair tracts formed during meiotic recombination in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (4) ◽  
pp. 1805-1814
Author(s):  
P Detloff ◽  
T D Petes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
High Frequency ◽  
Excision Repair ◽  
Yeast Saccharomyces Cerevisiae ◽  
Heteroduplex Dna ◽  
Homologous Chromosomes ◽  
His4 Gene

During meiotic recombination in the yeast Saccharomyces cerevisiae, heteroduplexes are formed at a high frequency between HIS4 genes located on homologous chromosomes. Using mutant alleles of the HIS4 gene that result in poorly repaired mismatches in heteroduplex DNA, we find that heteroduplexes often span a distance of 1.8 kb. In addition, we show that about one-third of the repair tracts initiated at well-repaired mismatches extend 900 bp.

scholarly journals Measurements of excision repair tracts formed during meiotic recombination in Saccharomyces cerevisiae.

1992 ◽  
Vol 12 (4) ◽  
pp. 1805-1814 ◽  
Author(s):  
P Detloff ◽  
T D Petes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
High Frequency ◽  
Excision Repair ◽  
Yeast Saccharomyces Cerevisiae ◽  
Heteroduplex Dna ◽  
Homologous Chromosomes ◽  
His4 Gene

During meiotic recombination in the yeast Saccharomyces cerevisiae, heteroduplexes are formed at a high frequency between HIS4 genes located on homologous chromosomes. Using mutant alleles of the HIS4 gene that result in poorly repaired mismatches in heteroduplex DNA, we find that heteroduplexes often span a distance of 1.8 kb. In addition, we show that about one-third of the repair tracts initiated at well-repaired mismatches extend 900 bp.

The Chromosome Bias of Misincorporations During Double-Strand Break Repair Is Not Altered in Mismatch Repair–Defective Strains of Saccharomyces cerevisiae

Genetics ◽  
1998 ◽  
Vol 148 (4) ◽  
pp. 1525-1533 ◽  
Author(s):  
Carolyn B McGill ◽  
Susan L Holbeck ◽  
Jeffrey N Strathern
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Mismatch Repair ◽  
Strand Break ◽  
Recombinational Repair ◽  
Dsb Repair ◽  
Site Specific ◽  
Initial Sequence ◽  
Dna Break ◽  
A Site

AbstractRecombinational repair of a site-specific, double-strand DNA break (DSB) results in increased reversion frequency for nearby mutations. Although some models for DSB repair predict that newly synthesized DNA will be inherited equally by both the originally broken chromosome and the chromosome that served as a template, the DNA synthesis errors are almost exclusively found on the chromosome that had the original DSB (introduced by the HO endonuclease). To determine whether mismatch repair acts on the template chromosome in a directed fashion to restore mismatches to the initial sequence, these experiments were repeated in mismatch repair-defective (pms1, mlh1, and msh2) backgrounds. The results suggest that mismatch repair is not responsible for the observed bias.

scholarly journals A 140-bp-Long Palindromic Sequence Induces Double-Strand Breaks During Meiosis in the Yeast Saccharomyces cerevisiae

Genetics ◽  
1997 ◽  
Vol 146 (3) ◽  
pp. 835-847 ◽  
Author(s):  
Dilip K Nag ◽  
Alicia Kurst
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Meiotic Recombination ◽  
Strand Break ◽  
Double Strand Break ◽  
Hairpin Structure ◽  
Palindromic Sequence ◽  
Yeast Saccharomyces Cerevisiae ◽  
Short Hairpin ◽  
Palindromic Sequences

Palindromic sequences have the potential to form hairpin or cruciform structures, which are putative substrates for several nucleases and mismatch repair enzymes. A genetic method was developed to detect such structures in vivo in the yeast Saccharomyces cerevisiae. Using this method we previously showed that short hairpin structures are poorly repaired by the mismatch repair system in S. cerevisiae. We show here that mismatches, when present in the stem of the hairpin structure, are not processed by the repair machinery, suggesting that they are treated differently than those in the interstrand base-paired duplex DNA. A 140-bp-long palindromic sequence, on the contrary, acts as a meiotic recombination hotspot by generating a site for a double-strand break, an initiator of meiotic recombination. We suggest that long palindromic sequences undergo cruciform extrusion more readily than short ones. This cruciform structure then acts as a substrate for structure-specific nucleases resulting in the formation of a double-strand break during meiosis in yeast. In addition, we show that residual repair of the short hairpin structure occurs in an MSH2-independent pathway.

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