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DNA unwinding heterogeneity by RecBCD results from static molecules able to equilibrate.

Liu, Bian Baskin, Ronald J Kowalczykowski, Stephen C

Published in Nature

Single-molecule studies can overcome the complications of asynchrony and ensemble-averaging in bulk-phase measurements, provide mechanistic insights into molecular activities, and reveal interesting variations between individual molecules. The application of these techniques to the RecBCD helicase of Escherichia coli has resolved some long-standing...

Pif1 family helicases suppress genome instability at G-quadruplex motifs

Paeschke, Katrin Bochman, Matthew L. Garcia, P. Daniela Cejka, Petr Friedman, Katherine L. Kowalczykowski, Stephen C. Zakian, Virginia A.

Published in Nature

Pif1 family DNA helicases are highly conserved, from bacteria to humans. Virginia Zakian and colleagues show that in yeast, human PIF1 can unwind a four-stranded structure known as a G-quadruplex. In doing so, it suppresses the genomic instability that arises at such structures. The ability of the human protein to complement in yeast demonstrates t...

Direct observation of individual RecA filaments assembling on single DNA molecules.

Galletto, R Ichiro Amitani Baskin, Rj Stephen Charles Kowalczykowski

Published in Nature

Escherichia coli RecA is essential for the repair of DNA double-strand breaks by homologous recombination. Repair requires the formation of a RecA nucleoprotein filament. Previous studies have indicated a mechanism of filament assembly whereby slow nucleation of RecA protein on DNA is followed by rapid growth. However, many aspects of this process ...

Structural biology: snapshots of DNA repair.

Stephen Charles Kowalczykowski

Published in Nature

The Srs2 helicase prevents recombination by disrupting Rad51 nucleoprotein filaments.

Veaute, X Jeusset, J Soustelle, C Stephen Charles Kowalczykowski Le Cam, E Fabre, F

Published in Nature

Homologous recombination is a ubiquitous process with key functions in meiotic and vegetative cells for the repair of DNA breaks. It is initiated by the formation of single-stranded DNA on which recombination proteins bind to form a nucleoprotein filament that is active in searching for homology, in the formation of joint molecules and in the excha...

Crystal structure of RecBCD enzyme reveals a machine for processing DNA breaks.

Singleton, Mr Dillingham, Ms Gaudier, M Stephen Charles Kowalczykowski Wigley, Db

Published in Nature

RecBCD is a multi-functional enzyme complex that processes DNA ends resulting from a double-strand break. RecBCD is a bipolar helicase that splits the duplex into its component strands and digests them until encountering a recombinational hotspot (Chi site). The nuclease activity is then attenuated and RecBCD loads RecA onto the 3 tail of the DNA....

Direct imaging of RecA nucleation and growth on single molecules of SSB-coated ssDNA.

Bell, Jason C Plank, Jody L Dombrowski, Christopher C Kowalczykowski, Stephen C

Published in Nature

Escherichia coli RecA is the defining member of a ubiquitous class of DNA strand-exchange proteins that are essential for homologous recombination, a pathway that maintains genomic integrity by repairing broken DNA. To function, filaments of RecA must nucleate and grow on single-stranded DNA (ssDNA) in direct competition with ssDNA-binding protein ...

Purified human BRCA2 stimulates RAD51-mediated recombination.

Jensen, Rb Carreira, A Stephen Charles Kowalczykowski

Published in Nature

Mutation of the breast cancer susceptibility gene, BRCA2, leads to breast and ovarian cancers. Mechanistic insight into the functions of human BRCA2 has been limited by the difficulty of isolating this large protein (3,418 amino acids). Here we report the purification of full-length BRCA2 and show that it both binds RAD51 and potentiates recombinat...

Single-molecule imaging of DNA pairing by RecA reveals a three-dimensional homology search.

Forget, Al Stephen Charles Kowalczykowski

Published in Nature

DNA breaks can be repaired with high fidelity by homologous recombination. A ubiquitous protein that is essential for this DNA template-directed repair is RecA. After resection of broken DNA to produce single-stranded DNA (ssDNA), RecA assembles on this ssDNA into a filament with the unique capacity to search and find DNA sequences in double-strand...

Cancer: catalyst of a catalyst.

Stephen Charles Kowalczykowski

Published in Nature

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