Cleavage of Model Replication Forks by Fission Yeast Mus81-Eme1 and Budding Yeast Mus81-Mms4

The blockage of replication forks can result in the disassembly of the replicative apparatus and reversal of the fork to form a DNA junction that must be processed in order for replication to restart and sister chromatids to segregate at mitosis. Fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4...

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Bibliographic Details
Published in:The Journal of biological chemistry 2003-02, Vol.278 (9), p.6928-6935
Main Authors: Whitby, Matthew C, Osman, Fekret, Dixon, Julie
Format: Article
Language:English
Subjects:
Dimerization
DNA - biosynthesis
DNA - metabolism
DNA-Binding Proteins - genetics
Endonucleases - genetics
Escherichia coli - metabolism
Flap Endonucleases
Models, Genetic
Plasmids - metabolism
Protein Binding
Saccharomyces cerevisiae Proteins
Saccharomycetales - genetics
Saccharomycetales - metabolism
Schizosaccharomyces - genetics
Schizosaccharomyces - metabolism
Schizosaccharomyces pombe Proteins - genetics
Time Factors
Trans-Activators - genetics
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Summary:The blockage of replication forks can result in the disassembly of the replicative apparatus and reversal of the fork to form a DNA junction that must be processed in order for replication to restart and sister chromatids to segregate at mitosis. Fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4 are endonucleases that have been implicated in the processing of aberrant DNA junctions formed at stalled replication forks. Here we have investigated the activity of purified Mus81-Eme1 and Mus81-Mms4 on substrates that resemble DNA junctions that are expected to form when a replication fork reverses. Both enzymes cleave Holliday junctions and substrates that resemble normal replication forks poorly or not at all. However, forks where the equivalents of either both the leading and lagging strands or just the lagging strand are juxtaposed at the junction point, or where either the leading or lagging strand has been unwound to produce a fork with a single-stranded tail, are cleaved well. Cleavage sites map predominantly between 3 and 6 bp 5′ of the junction point. For most substrates the leading strand template is cleaved. The sole exception is a fork with a 5′ single-stranded tail, which is cleaved in the lagging strand template.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M210006200