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PMID: 3282237 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mitotic sectored colonies: evidence of heteroduplex DNA formation during direct repeat recombination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 8 ·1988-04-00 ·Pages 2696-700

Ronne H, Rothstein R

Abstract

In yeast meiosis, ascosporal colonies are sometimes sectored for a marker--i.e., half the colony has one allele and half has the other. This is interpreted as replicative resolution of heteroduplex DNA (hDNA) formed as a recombination intermediate. We have looked for similar evidence of hDNA formation during mitotic recombination between two repeated sequences on the same chromosome. The two repeats, an ochre suppressor and a wild-type tRNA gene, are separated by plasmid DNA and the URA3 marker. Recombination between the repeats excises the URA3 gene and one copy of the repeat, leaving either the wild-type tRNA or the suppressor on the chromosome. A red/white color assay is used to distinguish between the two. We find that some colonies that have lost the URA3 gene are sectored for the suppressor. This suggests that hDNA is formed across the anticodon during the recombination event and then resolved by replication. The disruption of either of two genes involved in recombination and repair, RAD1 and RAD52, does not significantly alter the frequency of sectored colony formation during plasmid excision.

MeSH Terms
Canavanine/metabolism Culture Media DNA Repair DNA, Fungal/genetics Genes, Fungal Mitosis Orotic Acid/analogs & derivatives,metabolism RNA, Transfer/genetics Recombination, Genetic Repetitive Sequences, Nucleic Acid Saccharomyces cerevisiae/genetics Selection, Genetic Suppression, Genetic
Chemicals
Culture Media DNA, Fungal Canavanine Orotic Acid 5-fluoroorotic acid RNA, Transfer
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Ronne H
Department of Genetics and Development, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
Rothstein R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.8.2696
Published
1988-04-00
Pages
2696-700
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC280065
Subset
IM
Grants
NIGMS NIH HHS · GM 34587 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.8.2696 PMC Full Text View journal details
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