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PMID: 14617801 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

RNA interference inhibition of Mus81 reduces mitotic recombination in human cells.

Molecular biology of the cell ·Vol. 15 ·No. 2 ·2004-00-00 ·Pages 552-62

Blais V, Gao H, Elwell CA, Boddy MN, Gaillard PHL, Russell P, McGowan CH

Abstract

Mus81 is a highly conserved endonuclease with homology to the XPF subunit of the XPF-ERCC1 complex. In yeast Mus81 associates with a second subunit, Eme1 or Mms4, which is essential for endonuclease activity in vitro and for in vivo function. Human Mus81 binds to a homolog of fission yeast Eme1 in vitro and in vivo. We show that recombinant Mus81-Eme1 cleaves replication forks, 3' flap substrates, and Holliday junctions in vitro. By use of differentially tagged versions of Mus81 and Eme1, we find that Mus81 associates with Mus81 and that Eme1 associates with Eme1. Thus, complexes containing two or more Mus81-Eme1 units could function to coordinate substrate cleavage in vivo. Down-regulation of Mus81 by RNA interference reduces mitotic recombination in human somatic cells. The recombination defect is rescued by expression of a bacterial Holliday junction resolvase. These data provide direct evidence for a role of Mus81-Eme1 in mitotic recombination in higher eukaryotes and support the hypothesis that Mus81-Eme1 resolves Holliday junctions in vivo.

MeSH Terms
Amino Acid Sequence Cell Nucleolus/genetics,metabolism Cell Survival/physiology Cloning, Molecular DNA Damage/physiology DNA-Binding Proteins/genetics,metabolism Endonucleases/genetics,metabolism HeLa Cells Holliday Junction Resolvases/metabolism Humans Mitosis/genetics Molecular Sequence Data Protein Binding RNA Interference Recombination, Genetic/genetics Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins/genetics,metabolism
Chemicals
DNA-Binding Proteins Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins Eme1 protein, S pombe Endonucleases MUS81 protein, S cerevisiae MUS81 protein, human Holliday Junction Resolvases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Blais Veronique
Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Gao Hui
Elwell Cherilyn A
Boddy Michael N
Gaillard Pierre-Henri L
Russell Paul
McGowan Clare H
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-08-0580
Published
2004-00-00
Epub
2003-00-14
Pages
552-62
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC329235
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e03-08-0580 PMC Full Text View journal details
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