A Single-Molecule Approach to Visualize the Unwinding Activity of DNA Helicases View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2011-07-01

AUTHORS

Natalia Fili , Christopher P. Toseland , Mark S. Dillingham , Martin R. Webb , Justin E. Molloy

ABSTRACT

Almost all aspects of DNA metabolism involve separation of double-stranded DNA catalyzed by helicases. Observation and measurement of the dynamics of these events at the single-molecule level provide important mechanistic details of helicase activity and give the opportunity to probe aspects that are not revealed in bulk solution measurements. The assay, presented here, provides information about helicase unwinding rates and processivity. Visualization is achieved by using a fluorescent single-stranded DNA-binding protein (SSB), which allows the time course of individual DNA unwinding events to be observed using total internal reflection fluorescence microscopy. Observation of a prototypical helicase, Bacillus subtilis AddAB, shows that the unwinding process consists of bursts of unwinding activity, interspersed with periods of pausing. More... »

PAGES

193-214

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-61779-261-8_13

DOI

http://dx.doi.org/10.1007/978-1-61779-261-8_13

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1032580563

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/21809208


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