RAD52-INDEPENDENT MITOTIC GENE CONVERSION IN SACCHAROMYCES CEREVISIAE FREQUENTLY RESULTS IN CHROMOSOMAL LOSS

Genetics ◽  
1985 ◽  
Vol 111 (1) ◽  
pp. 7-22
Author(s):  
James E Haber ◽  
Mark Hearn
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Mitotic Recombination ◽  
Gene Conversion Event ◽  
Intragenic Recombination ◽  
Wild Type ◽  
Conversion Event ◽  
Chromosomal Loss ◽  
Mitotic Gene Conversion ◽  
Striking Effect

ABSTRACT We have examined spontaneous, interchromosomal mitotic recombination events between his4 alleles in both Rad+ and rad52 strains of Saccharomyces cerevisiae. In Rad+ strains, 74% of the His+ prototrophs resulted from gene conversion events without exchange of flanking markers. In diploids homozygous for the rad52-1 mutation, the frequency of His+ prototroph formation was less than 5% of the wild-type value, and more than 80% of the gene conversion events were accompanied by an exchange of flanking markers. Most of the rad52 intragenic recombination events arose by gene conversion accompanied by an exchange of flanking markers and not by a simple reciprocal exchange between the his4A and his4C alleles. There were also profound effects on the kinds of recombinant products that were recovered. The most striking effect was that RAD52-independent mitotic recombination frequently results in the loss of one of the two chromosomes participating in the gene conversion event.

scholarly journals Spontaneous mitotic recombination in yeast: the hyper-recombinational rem1 mutations are alleles of the RAD3 gene.

Genetics ◽  
1988 ◽  
Vol 119 (2) ◽  
pp. 289-301
Author(s):  
B A Montelone ◽  
M F Hoekstra ◽  
R E Malone
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Excision Repair ◽  
Mitotic Recombination ◽  
Crossing Over ◽  
Wild Type ◽  
Recombination Repair ◽  
Mitotic Gene Conversion ◽  
Type Gene ◽  
Repair Genes

Abstract The RAD3 gene of Saccharomyces cerevisiae is required for UV excision-repair and is essential for cell viability. We have identified the rem1 mutations (enhanced spontaneous mitotic recombination and mutation) of Saccharomyces cerevisiae as alleles of RAD3 by genetic mapping, complementation with the cloned wild-type gene, and DNA hybridization. The high levels of spontaneous mitotic gene conversion, crossing over, and mutation conferred upon cells by the rem1 mutations are distinct from the effects of all other alleles of RAD3. We present preliminary data on the localization of the rem1 mutations within the RAD3 gene. The interaction of the rem1 mutant alleles with a number of radiation-sensitive mutations is also different than the interactions reported for previously described (UV-sensitive) alleles of RAD3. Double mutants of rem1 and a defect in the recombination-repair pathway are inviable, while double mutants containing UV-sensitive alleles of RAD3 are viable. The data presented here demonstrate that: (1) rem1 strains containing additional mutations in other excision-repair genes do not exhibit elevated gene conversion; (2) triple mutants containing rem1 and mutations in both excision-repair and recombination-repair are viable; (3) such triple mutants containing rad52 have reduced levels of gene conversion but wild-type frequencies of crossing over. We have interpreted these observations in a model to explain the effects of rem1. Consistent with the predictions of the model, we find that the size of DNA from rem1 strains, as measured by neutral sucrose gradients, is smaller than wild type.

Homothallic switching of Saccharomyces cerevisiae mating type genes by using a donor containing a large internal deletion

1985 ◽  
Vol 5 (8) ◽  
pp. 2154-2158
Author(s):  
B Weiffenbach ◽  
J E Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Mating Type ◽  
Gene Conversion Event ◽  
Conversion Event ◽  
Base Pair Deletion ◽  
Mating Type Genes ◽  
Type Gene ◽  
Mat Locus ◽  
Donor Locus

Homothallic switching of the mating type genes of Saccharomyces cerevisiae occurs by a gene conversion event, replacing sequences at the expressed MAT locus with a DNA segment copied from one of two unexpressed loci, HML or HMR. The transposed Ya or Y alpha sequences are flanked by homologous regions that are believed to be essential for switching. We examined the transposition of a mating type gene (hmr alpha 1-delta 6) which contains a 150-base-pair deletion spanning the site where the HO endonuclease generates a double-stranded break in MAT that initiates the gene conversion event. Despite the fact that the ends of the cut MAT region no longer share homology with the donor hmr alpha 1-delta 6, switching of MATa or MAT alpha to mat alpha 1-delta 6 was efficient. However, there was a marked increase in the number of aberrant events, especially the formation of haploid-inviable fusions between MAT and the hmr alpha 1-delta 6 donor locus.

scholarly journals Homothallic switching of Saccharomyces cerevisiae mating type genes by using a donor containing a large internal deletion.

1985 ◽  
Vol 5 (8) ◽  
pp. 2154-2158 ◽  
Author(s):  
B Weiffenbach ◽  
J E Haber
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Mating Type ◽  
Gene Conversion Event ◽  
Conversion Event ◽  
Base Pair Deletion ◽  
Mating Type Genes ◽  
Type Gene ◽  
Mat Locus ◽  
Donor Locus

Homothallic switching of the mating type genes of Saccharomyces cerevisiae occurs by a gene conversion event, replacing sequences at the expressed MAT locus with a DNA segment copied from one of two unexpressed loci, HML or HMR. The transposed Ya or Y alpha sequences are flanked by homologous regions that are believed to be essential for switching. We examined the transposition of a mating type gene (hmr alpha 1-delta 6) which contains a 150-base-pair deletion spanning the site where the HO endonuclease generates a double-stranded break in MAT that initiates the gene conversion event. Despite the fact that the ends of the cut MAT region no longer share homology with the donor hmr alpha 1-delta 6, switching of MATa or MAT alpha to mat alpha 1-delta 6 was efficient. However, there was a marked increase in the number of aberrant events, especially the formation of haploid-inviable fusions between MAT and the hmr alpha 1-delta 6 donor locus.

scholarly journals Isolation and characterization of Schizosaccharomyces pombe mutants affected in mitotic recombination.

Genetics ◽  
1991 ◽  
Vol 128 (3) ◽  
pp. 495-504
Author(s):  
A Gysler-Junker ◽  
Z Bodi ◽  
J Kohli
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Mitotic Recombination ◽  
Selective Medium ◽  
Intragenic Recombination ◽  
Crossing Over ◽  
Wild Type ◽  
Isolation And Characterization ◽  
Mitotic Gene Conversion ◽  
Type Frequency

Abstract A haploid Schizosaccharomyces pombe strain carrying a heteroallelic duplication of the ade6 gene was used to isolate mitotic recombination-deficient mutants. Recombination between the different copies of the ade6 gene can lead to Ade+ segregants. These are observed as growing papillae when colonies of a suitable size are replicated onto selective medium. We isolated mutants which show an altered papillation phenotype. With two exceptions, they exhibit a decrease in the frequency of mitotic recombination between the heteroalleles of the duplication. The two other mutants display a hyper-recombination phenotype. The 12 mutations were allocated to at least nine distinct loci by recombination tests. Of the eight rec mutants analyzed further, six were also affected in mitotic intergenic recombination in the intervals cen2-mat or cen3-arg 1. No effect on mitotic intragenic recombination was observed. These data suggest that mitotic gene conversion and crossing over can be separated mutationally. Meiotic recombination occurs at the wild-type frequency in all mutants investigated.

scholarly journals Studies on mitotic gene conversion in Ustilago

1966 ◽  
Vol 8 (3) ◽  
pp. 323-337 ◽  
Author(s):  
Robin Holliday
Keyword(s):  
Gene Conversion ◽  
Mitotic Cycle ◽  
Uv Light ◽  
Reductase Activity ◽  
Sole Source ◽  
Intragenic Recombination ◽  
Wild Type ◽  
Back Mutation ◽  
Mitotic Gene Conversion ◽  
Hybrid Dna

In order to develop a system for the study of the mechanism of intragenic recombination in Ustilago, mutants lacking nitrate reductase activity were isolated, and five alleles were combined in pairs in ten vegetative heteroallelic diploids. The diploids have the mutant phenotype, i.e. inability to utilize nitrate as sole source of nitrogen, but they will recombine to produce wild-type cells much more frequently than the back-mutation rates of haploids or homoallelic diploids. The spontaneous rate of recombination can be enormously increased by low doses of UV light, particularly if treatment is during the period of DNA synthesis in the mitotic cycle. By means of half-tetrad analysis it has been shown that this process of intragenic recombination, as in other fungi, is due to gene conversion rather than reciprocal exchange. It has also been shown that the frequency of UV-induced conversion under standard conditions gives a rough measure of the distance between two mutant sites, since it was possible to use these frequencies to make a linear fine structure map of the gene. These results are discussed in relation to a hybrid DNA model for gene conversion slightly modified from that previously suggested for meiotic recombination.

scholarly journals Stimulation of Mitotic Recombination Events by High Levels of RNA Polymerase II Transcription in Yeast

2000 ◽  
Vol 20 (15) ◽  
pp. 5404-5414 ◽  
Author(s):  
Dean Saxe ◽  
Abhijit Datta ◽  
Sue Jinks-Robertson
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Gene Conversion ◽  
Regulatory Protein ◽  
Mitotic Recombination ◽  
Gene Conversion Event ◽  
Conversion Event ◽  
The Impact ◽  
Protein Transcription ◽  
Rna Polymerase Ii Transcription

ABSTRACT The impact of high levels of RNA polymerase II transcription on mitotic recombination was examined using lys2 recombination substrates positioned on nonhomologous chromosomes. Substrates were used that could produce Lys+ recombinants by either a simple (noncrossover) gene conversion event or a crossover-associated recombination event, by only a simple gene conversion event, or by only a crossover event. Transcription of the lys2 substrates was regulated by the highly inducible GAL1-10 promoter or the low-level LYS2 promoter, with GAL1-10 promoter activity being controlled by the presence or absence of the Gal80p negative regulatory protein. Transcription was found to stimulate recombination in all assays used, but the level of stimulation varied depending on whether only one or both substrates were highly transcribed. In addition, there was an asymmetry in the types of recombination events observed when one substrate versus the other was highly transcribed. Finally, the lys2 substrates were positioned as direct repeats on the same chromosome and were found to exhibit a different recombinational response to high levels of transcription from that exhibited by the repeats on nonhomologous chromosomes. The relevance of these results to the mechanisms of transcription-associated recombination are discussed.

scholarly journals Characterization of the Roles of the Saccharomyces cerevisiae RAD54 Gene and a Homologue of RAD54, RDH54/TID1, in Mitosis and Meiosis

Genetics ◽  
1997 ◽  
Vol 147 (4) ◽  
pp. 1545-1556 ◽  
Author(s):  
Miki Shinohara ◽  
Emi Shita-Yamaguchi ◽  
Jean-Marie Buerstedde ◽  
Hideo Shinagawa ◽  
Hideyuki Ogawa ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Double Mutant ◽  
Mitotic Recombination ◽  
Genome Project ◽  
Yeast Genome ◽  
Wild Type ◽  
Reduced Frequency ◽  
A Minor ◽  
Mitosis And Meiosis

Abstract The RAD54 gene, which encodes a protein in the SW12/SNF2 family, plays an important role in recombination and DNA repair in Saccharomyces cerevisiae. The yeast genome project revealed a homologue of RAD54, RDH54/TID1. Properties of the rdh54/tid1 mutant and the rad54 rdh54/tid1 double mutant are shown for mitosis and meiosis. The rad54 mutant is sensitive to the alkylating agent, methyl methanesulfonate (MMS), and is defective in interchromosomal and intrachromosomal gene conversion. The rdh54/tid1 single mutant, on the other hand, does not show any significant deficiency in mitosis. However, the rad54 rdh54/tid1 mutant is more sensitive to MMS and more defective in interchromosomal gene conversion than is the rad54 mutant, but shows the same frequency of intrachromosomal gene conversion as the rad54 mutant. These results suggest that RDH54/TID1 is involved in a minor pathway of mitotic recombination in the absence of RAD54. In meiosis, both single mutants produce viable spores at slightly reduced frequency. However, only the rdh54/tid1 mutant, but not the rad54 mutant, shows significant defects in recombination: retardation of the repair of meiosis-specific double-strand breaks (DSBs) and delayed formation of physical recombinants. Furthermore, the rad54 rdh54/tid1 double mutant is completely defective in meiosis, accumulating DSBs with more recessed ends than the wild type and producing fewer physical recombinants than the wild type. These results suggest that one of the differences between the late stages of mitotic recombination and meiotic recombination might be specified by differential dependency on the Rad54 and Rdh54/Tid1 proteins.

The Escherichia coli recA gene increases UV-induced mitotic gene conversion in Saccharomyces cerevisiae

1994 ◽  
Vol 25 (5) ◽  
pp. 472-474 ◽  
Author(s):  
Viera Vlčková ◽  
Luba Černáková ◽  
Eva Farkašová ◽  
Jela Brozmanová
Keyword(s):  
Escherichia Coli ◽  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Reca Gene ◽  
Mitotic Gene Conversion

Evidence for Independent Mismatch Repair Processing on Opposite Sides of a Double-Strand Break in Saccharomyces cerevisiae

Genetics ◽  
1998 ◽  
Vol 148 (1) ◽  
pp. 59-70
Author(s):  
Yi-shin Weng ◽  
Jac A Nickoloff
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Conversion ◽  
Mismatch Repair ◽  
Strand Break ◽  
Double Strand Break ◽  
Stem Loop ◽  
Mitotic Gene Conversion ◽  
Low Levels ◽  
Mat Locus ◽  
Insertion Mutations

Abstract Double-strand break (DSB) induced gene conversion in Saccharomyces cerevisiae during meiosis and MAT switching is mediated primarily by mismatch repair of heteroduplex DNA (hDNA). We used nontandem ura3 duplications containing palindromic frameshift insertion mutations near an HO nuclease recognition site to test whether mismatch repair also mediates DSB-induced mitotic gene conversion at a non-MAT locus. Palindromic insertions included in hDNA are expected to produce a stem-loop mismatch, escape repair, and segregate to produce a sectored (Ura+/−) colony. If conversion occurs by gap repair, the insertion should be removed on both strands, and converted colonies will not be sectored. For both a 14-bp palindrome, and a 37-bp near-palindrome, ~75% of recombinant colonies were sectored, indicating that most DSB-induced mitotic gene conversion involves mismatch repair of hDNA. We also investigated mismatch repair of well-repaired markers flanking an unrepaired palindrome. As seen in previous studies, these additional markers increased loop repair (likely reflecting corepair). Among sectored products, few had additional segregating markers, indicating that the lack of repair at one marker is not associated with inefficient repair at nearby markers. Clear evidence was obtained for low levels of short tract mismatch repair. As seen with full gene conversions, donor alleles in sectored products were not altered. Markers on the same side of the DSB as the palindrome were involved in hDNA less often among sectored products than nonsectored products, but markers on the opposite side of the DSB showed similar hDNA involvement among both product classes. These results can be explained in terms of corepair, and they suggest that mismatch repair on opposite sides of a DSB involves distinct repair tracts.

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