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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Main Authors: Whitby, M, Osman, F, Dixon, J
Format: Journal article
Language:English
Published: 2003
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author Whitby, M
Osman, F
Dixon, J
author_facet Whitby, M
Osman, F
Dixon, J
author_sort Whitby, M
collection OXFORD
description 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.
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spelling oxford-uuid:feba8563-193d-4794-b99d-714d8d5434672022-03-27T13:38:49ZCleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:feba8563-193d-4794-b99d-714d8d543467EnglishSymplectic Elements at Oxford2003Whitby, MOsman, FDixon, JThe 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.
spellingShingle Whitby, M
Osman, F
Dixon, J
Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title_full Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title_fullStr Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title_full_unstemmed Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title_short Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
title_sort cleavage of model replication forks by fission yeast mus81 eme1 and budding yeast mus81 mms4
work_keys_str_mv AT whitbym cleavageofmodelreplicationforksbyfissionyeastmus81eme1andbuddingyeastmus81mms4
AT osmanf cleavageofmodelreplicationforksbyfissionyeastmus81eme1andbuddingyeastmus81mms4
AT dixonj cleavageofmodelreplicationforksbyfissionyeastmus81eme1andbuddingyeastmus81mms4