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PMID: 8929389 Published · ppublish English Journal Article

Stimulation of mitotic recombination upon transcription from the yeast GAL1 promoter but not from other RNA polymerase I, II and III promoters.

Current genetics ·Vol. 30 ·No. 5 ·1996-11-00 ·Pages 381-8

Bratty J, Ferbeyre G, Molinaro C, Cedergren R

Abstract

Homologous recombination in Saccharomyces cerevisiae and other organisms can be stimulated by transcription. Consistent with this, we find that recombination of a chromosomal ade1 allele with a plasmid-borne ADE1 ORF under the control of the GAL1 promoter increased from 6.1x10(-6) to 1.7x10(-4) when transcription of the plasmid locus was induced by growing the cells in the presence of galactose. Recombination could also be stimulated by over-expressing the Gal4 transcription factor in the presence of the GAL1-ADE1 plasmid, while culturing the cells in dextrose medium. However, when transcription of the same ORF was driven from the highly active promoters of the rDNA (RNA polymerase I), and ADH1 (RNA polymerase II) genes, only background levels of recombination (5-10x10(-6)) were observed, irrespective of the carbon source. Recombination was found to involve integration of the whole plasmid and to depend on RAD51, RAD52 and RAD54. The results indicate that increased accessibility of transcriptionally active chromatin is not sufficient to cause increased rates of this kind of reciprocal exchange.

MeSH Terms
Alleles Blotting, Northern Blotting, Southern Chromosome Mapping DNA Helicases DNA Repair Enzymes DNA, Recombinant/physiology DNA-Binding Proteins/genetics Fungal Proteins/genetics Galactose/genetics Gene Expression Regulation, Fungal Models, Genetic Open Reading Frames Plasmids/genetics Polymerase Chain Reaction Promoter Regions, Genetic RNA Polymerase I/genetics RNA Polymerase II/genetics RNA Polymerase III/genetics Rad51 Recombinase Rad52 DNA Repair and Recombination Protein Recombination, Genetic Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Transcription Factors Transcription, Genetic
Chemicals
DNA, Recombinant DNA-Binding Proteins Fungal Proteins GAL4 protein, S cerevisiae RAD52 protein, S cerevisiae Rad52 DNA Repair and Recombination Protein Saccharomyces cerevisiae Proteins Transcription Factors RAD51 protein, S cerevisiae RNA Polymerase II Rad51 Recombinase RNA Polymerase I RNA Polymerase III RAD54 protein, S cerevisiae DNA Helicases DNA Repair Enzymes Galactose
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bratty J
Département de biochimie, Université de Montréal, Montréal, H3C 3J7, Canada.
Ferbeyre G
Molinaro C
Cedergren R
Article Info
Journal
Current genetics
Abbr.
Curr Genet
ISSN
0172-8083
Published
1996-11-00
Pages
381-8
Language
English
Region
United States
NLM ID
8004904
Subset
IM
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