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PMID: 10835372 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Partial suppression of the fission yeast rqh1(-) phenotype by expression of a bacterial Holliday junction resolvase.

The EMBO journal ·Vol. 19 ·No. 11 ·2000-06-01 ·Pages 2751-62

Doe CL, Dixon J, Osman F, Whitby MC

Abstract

A key stage during homologous recombination is the processing of the Holliday junction, which determines the outcome of the recombination reaction. To dissect the pathways of Holliday junction processing in a eukaryote, we have targeted an Escherichia coli Holliday junction resolvase to the nuclei of fission yeast recombination-deficient mutants and analysed their phenotypes. The resolvase partially complements the UV and hydroxyurea hypersensitivity and associated aberrant mitoses of an rqh1(-) mutant. Rqh1 is a member of the RecQ subfamily of DNA helicases that control recombination particularly during S-phase. Significantly, overexpression of the resolvase in wild-type cells partly mimics the loss of viability, hyper-recombination and 'cut' phenotype of an rqh1(-) mutant. These results indicate that Holliday junctions form in wild-type cells that are normally removed in a non-recombinogenic way, possibly by Rqh1 catalysing their reverse branch migration. We propose that in the absence of Rqh1, replication fork arrest results in the accumulation of Holliday junctions, which can either impede sister chromatid segregation or lead to the formation of recombinants through Holliday junction resolution.

MeSH Terms
Cell Nucleus/metabolism DNA Helicases/deficiency,genetics DNA, Fungal/genetics,metabolism Endodeoxyribonucleases/genetics,physiology Escherichia coli/enzymology Escherichia coli Proteins Eukaryotic Cells/metabolism Fungal Proteins/genetics,physiology Genes, Reporter Genes, Synthetic Genetic Complementation Test Green Fluorescent Proteins Holliday Junction Resolvases Luminescent Proteins/genetics Models, Genetic Nuclear Localization Signals Nuclear Proteins/genetics,physiology Phenotype Radiation Tolerance/genetics Recombinant Fusion Proteins/physiology Recombination, Genetic/physiology S Phase Schizosaccharomyces/genetics,metabolism,radiation effects Schizosaccharomyces pombe Proteins Ultraviolet Rays
Chemicals
DNA, Fungal Escherichia coli Proteins Fungal Proteins Luminescent Proteins Nuclear Localization Signals Nuclear Proteins Recombinant Fusion Proteins Schizosaccharomyces pombe Proteins Green Fluorescent Proteins Endodeoxyribonucleases Holliday Junction Resolvases RusA protein, E coli DNA Helicases Rqh1 protein, S pombe
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Doe C L
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Dixon J
Osman F
Whitby M C
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.11.2751
Published
2000-06-01
Pages
2751-62
Language
English
Region
England
NLM ID
8208664
PMCID
PMC212752
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/19.11.2751 PMC Full Text View journal details
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