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BLM helicase measures DNA unwound before switching strands and hRPA promotes unwinding reinitiation
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SYSNO ASEP 0333639 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title BLM helicase measures DNA unwound before switching strands and hRPA promotes unwinding reinitiation Author(s) Yodh, J.G. (US)
Stevens, B.C. (US)
Kanagaraj, R. (CH)
Janščák, Pavel (UMG-J) RID
Ha, T. (US)Number of authors 5 Source Title EMBO Journal. - : Wiley - ISSN 0261-4189
Roč. 28, č. 4 (2009), s. 405-416Number of pages 12 s. Language eng - English Country GB - United Kingdom Keywords Bloom syndrome ; FRET ; helicase ; hRPA ; single molecule Subject RIV EB - Genetics ; Molecular Biology CEZ AV0Z50520514 - UMG-J (2005-2011) UT WOS 000263556200012 DOI 10.1038/emboj.2008.298 Annotation Bloom syndrome (BS) is characterized by genomic instability and high predisposition to cancer. BLM, gene defective in BS, encodes a member of the RecQ family of 3´-5´ DNA helicases and is proposed to function in recombinational repair at DNA replication. We used single-molecule fluorescence resonance energy transfer microscopy to examine BLM behaviour on forked DNA substrates. BLM unwound individual DNA molecules repetitively, a short length unwound followed by rapid reannealing in successions. We show that monomeric BLM can "measure" how many base pairs it has unwound, and after a critical length it reverses the unwinding through strand switching and translocating on the opposing strand. Repetitive unwinding persisted even in the presence of hRPA, and interaction between wild-type BLM and hRPA was necessary for unwinding reinitiation on hRPA-coated DNA. These activities may facilitate BLM processing of stalled replication forks and illegitimately formed recombination intermediates. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2010
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