Fbh1 limits Rad51-dependent recombination at blocked replication forks

Alexander Lorenz, Fekret Osman, Victoria Folkyte, Sevil Sofueva, Matthew C. Whitby

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Controlling the loading of Rad51 onto DNA is important for governing when and how homologous recombination is used. Here we use a combination of genetic assays and indirect immunofluorescence to show that the F-box DNA helicase (Fbh1) functions in direct opposition to the Rad52 orthologue Rad22 to curb Rad51 loading onto DNA in fission yeast. Surprisingly, this activity is unnecessary for limiting spontaneous directrepeat recombination. Instead it appears to play an important role in preventing recombination when replication forks are blocked and/or broken. When overexpressed, Fbh1 specifically reduces replication fork block-induced recombination, as well as the number of Rad51 nuclear foci that are induced by replicative stress. These abilities are dependent on its DNA helicase/translocase activity, suggesting that Fbh1 exerts its control on recombination by acting as a Rad51 disruptase. In accord with this, overexpression of Fbh1 also suppresses the high levels of recombinant formation and Rad51 accumulation at a site-specific replication fork barrier in a strain lacking the Rad51 disruptase Srs2. Similarly overexpression of Srs2 suppresses replication fork block-induced gene conversion events in an fbh1Δ mutant, although an inability to suppress deletion events suggests that Fbh1 has a distinct functionality, which is not readily substituted by Srs2.
Original languageEnglish
Pages (from-to)4742-4756
Number of pages15
JournalMolecular and Cellular Biology
Volume29
Issue number17
Early online date22 Jun 2009
DOIs
Publication statusPublished - Sep 2009

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Genetic Recombination
DNA Helicases
Gene Conversion
Schizosaccharomyces
Homologous Recombination
DNA
Indirect Fluorescent Antibody Technique

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Fbh1 limits Rad51-dependent recombination at blocked replication forks. / Lorenz, Alexander; Osman, Fekret; Folkyte, Victoria; Sofueva, Sevil; Whitby, Matthew C.

In: Molecular and Cellular Biology, Vol. 29, No. 17, 09.2009, p. 4742-4756.

Research output: Contribution to journalArticle

Lorenz, Alexander ; Osman, Fekret ; Folkyte, Victoria ; Sofueva, Sevil ; Whitby, Matthew C. / Fbh1 limits Rad51-dependent recombination at blocked replication forks. In: Molecular and Cellular Biology. 2009 ; Vol. 29, No. 17. pp. 4742-4756.
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