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Real-time observation of RecA filament nucleation and dynamics

Joo, Chirlmin and Mckinney, Sean and Cissé, Ibrahim and Okumus, Burak and Nakamura, Muneaki and Rasnik, Ivan and Myong, Sua and Ha, Taekjip (2007) Real-time observation of RecA filament nucleation and dynamics. In: 234th ACS National Meeting & Exposition, 19-23 August 2007, Boston, MA. https://resolver.caltech.edu/CaltechAUTHORS:20201214-074214212

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Abstract

The RecA protein helps maintain genomic integrity through recombination. Using new single-mol. fluorescence assays and hidden Markov modeling, we first show the most direct evidence that a RecA filament grows and shrinks primarily one monomer at a time and only at the extremities. Both ends grow and shrink, contrary to expectation, but a higher binding rate at one end is responsible for directional filament growth. Next, we est. that about five monomers are sufficient for filament nucleation. We further find that the nucleation is likely to occur through the binding of a preassembled oligomer rather than the simultaneous binding of several monomers, using a vesicle encapsulation assay. Finally, although ordinarily SSB prevents filament nucleation, single RecA monomers can easily be added to an existing filament and displace SSB from DNA at the rate of filament extension. This supports the proposal for a passive role of RecA-loading machineries in SSB removal.


Item Type:Conference or Workshop Item (Paper)
ORCID:
AuthorORCID
Joo, Chirlmin0000-0003-2803-0335
Cissé, Ibrahim0000-0002-8764-1809
Okumus, Burak0000-0001-9017-8467
Ha, Taekjip0000-0003-2195-6258
Additional Information:© 2007 American Chemical Society.
Record Number:CaltechAUTHORS:20201214-074214212
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201214-074214212
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107053
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Dec 2020 21:12
Last Modified:14 Dec 2020 21:12

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