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PMID: 8355651 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Consequences of unique double-stranded breaks in yeast chromosomes: death or homozygosis.

Molecular & general genetics : MGG ·Vol. 240 ·No. 2 ·1993-08-00 ·Pages 170-8

Fairhead C, Dujon B

Abstract

We have developed a system in which a unique double-stranded break (DSB) can be introduced into a yeast chromosome during mitotic growth. The recognition site for the endonuclease I-SceI was inserted at different places in the yeast genome in haploid and diploid cells expressing this endonuclease. Induction of the break in haploids results in cell death if no intact copy of the cleaved region is present in the cell. If such a copy is provided on a plasmid, as an ectopic gene duplication, or on a homologous chromosome, the break can be repaired. Repair results in two identical copies in the genome of the locus which has been cut. We call this phenomenon homozygotization by reference to diploids heterozygous for the cut site in which repair leads to homozygosis at this site. We have compared the efficiencies of repair in the various topological situations examined, and conclude that some mechanism must search for regions of homology to both sides of the DSB and that repair is successful only if the homologies are provided by the same template molecule.

MeSH Terms
Chromosomes, Fungal Cloning, Molecular DNA Damage DNA Repair DNA, Fungal Deoxyribonucleases, Type II Site-Specific/metabolism Diploidy Electrophoresis, Gel, Pulsed-Field Fungi/genetics,growth & development Galactose/metabolism Haploidy Heterozygote Homozygote Restriction Mapping Saccharomyces cerevisiae Proteins
Chemicals
DNA, Fungal Saccharomyces cerevisiae Proteins SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific Galactose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fairhead C
Unité de Génétique moléculaire des Levures (URA 1149 du CNRS), Institut Pasteur, Paris, France.
Dujon B
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00277054
Published
1993-08-00
Pages
170-8
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00277054
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