Dynamic look at DNA unwinding by a replicative helicase

Lee, S. J., Syed, S., Enemark, E. J., Schuck, S., Stenlund, A., Ha, T., Joshua-Tor, L. (March 2014) Dynamic look at DNA unwinding by a replicative helicase. Proceedings of the National Academy of Sciences of the United States of America, 111 (9). E827-35. ISSN 0027-8424

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URL: http://www.ncbi.nlm.nih.gov/pubmed/24550505
DOI: 10.1073/pnas.1322254111

Abstract

A prerequisite for DNA replication is the unwinding of duplex DNA catalyzed by a replicative hexameric helicase. Despite a growing body of research, key elements of helicase mechanism remain under substantial debate. In particular, the number of DNA strands encircled by the helicase ring during unwinding and the ring orientation at the replication fork completely contrast in contemporary mechanistic models. Here we use single-molecule and ensemble assays to address these questions for the papillomavirus E1 helicase. We find that E1 unwinds DNA with a strand-exclusion mechanism, with the N-terminal side of the helicase ring facing the replication fork. We show that E1 generates strikingly heterogeneous unwinding patterns stemming from varying degrees of repetitive movements, which is modulated by the DNA-binding domain. Together, our studies reveal previously unrecognized dynamic facets of replicative helicase unwinding mechanisms.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > helicase
CSHL Authors:
Communities: CSHL labs > Joshua-Tor lab
CSHL labs > Stenlund lab
CSHL Cancer Center Program > Gene Regulation and Cell Proliferation
Depositing User: Matt Covey
Date: 4 March 2014
Date Deposited: 07 Mar 2014 16:47
Last Modified: 20 Dec 2017 21:53
PMCID: PMC3948270
Related URLs:
URI: https://repository.cshl.edu/id/eprint/29569

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