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RecQ helicase and topoisomerase III comprise a novel DNA strand passage function: a conserved mechanism for control of D...

Harmon, Fg Digate, Rj Stephen Charles Kowalczykowski

Published in Molecular Cell

E. coli RecQ protein is a multifunctional helicase with homologs that include the S. cerevisiae Sgs1 helicase and the H. sapiens Wrn and Blm helicases. Here we show that RecQ helicase unwinds a covalently closed double-stranded DNA (dsDNA) substrate and that this activity specifically stimulates E. coli topoisomerase III (Topo III) to fully catenat...

Decatenation of DNA by the S. cerevisiae Sgs1-Top3-Rmi1 and RPA complex: a mechanism for disentangling chromosomes.

Cejka, Petr Plank, Jody L Dombrowski, Christopher C Kowalczykowski, Stephen C

Published in Molecular cell

Genetic evidence indicates that Saccharomyces cerevisiae Sgs1, Top3, and Rmi1 resolve topologically linked intermediates arising from DNA replication and recombination. Using purified proteins, we show that Sgs1, Top3, Rmi1, and replication protein A (RPA) coordinate catenation and decatenation of dsDNA through sequential passage of single strands ...

RecFOR proteins load RecA protein onto gapped DNA to accelerate DNA strand exchange: a universal step of recombinational...

Katsumi Morimatsu Stephen Charles Kowalczykowski

Published in Molecular Cell

Genetic evidence suggests that the RecF, RecO, and RecR (RecFOR) proteins participate in a common step of DNA recombination and repair, yet the biochemical event requiring collaboration of all three proteins is unknown. Here, we show that the concerted action of the RecFOR complex directs the loading of RecA protein specifically onto gapped DNA tha...

A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Re...

Wang, At Taeho Kim Wagner, Je Conti, Ba Lach, Fp Huang, Al Molina, H Sanborn, Em Zierhut, H Cornes, Bk ...

Published in Molecular Cell

Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. In vitro, RAD51-T131P displays DNA-independent AT...

The RecA binding locus of RecBCD is a general domain for recruitment of DNA strand exchange proteins.

Spies, M Stephen Charles Kowalczykowski

Published in Molecular Cell

RecBCD enzyme facilitates loading of RecA protein onto ssDNA produced by its helicase/nuclease activity. This process is essential for RecBCD-mediated homologous recombination. Here, we establish that the C-terminal nuclease domain of the RecB subunit (RecBnuc) forms stable complexes with RecA. Interestingly, RecBnuc also interacts with and loads n...

Top3-Rmi1 dissolve Rad51-mediated D loops by a topoisomerase-based mechanism.

Fasching, Cl Cejka, P Stephen Charles Kowalczykowski Heyer, Wd

Published in Molecular Cell

The displacement loop (D loop) is a DNA strand invasion product formed during homologous recombination. Disruption of nascent D loops prevents recombination, and during synthesis-dependent strand annealing (SDSA), disruption of D loops extended by DNA polymerase ensures a non-crossover outcome. The proteins implicated in D loop disruption are DNA m...

Visualization of Rad54, a chromatin remodeling protein, translocating on single DNA molecules.

Ichiro Amitani Baskin, Rj Stephen Charles Kowalczykowski

Published in Molecular Cell

Rad54 protein plays an important role in the recombinational repair of double-strand DNA (dsDNA) breaks. It is a dsDNA-dependent ATPase that belongs to the Swi2/Snf2 family of chromatin-remodeling proteins. Rad54 remodels (1) DNA structure, (2) chromatin structure, and (3) Rad51-dsDNA complexes. These abilities imply that Rad54 moves along DNA. Her...

A step backward in advancing DNA replication: rescue of stalled replication forks by RecG.

Dillingham, Ms Stephen Charles Kowalczykowski

Published in Molecular Cell

In the October 5 issue of Cell, Singleton et al. report the crystal structure of RecG protein bound to an analog of a stalled DNA replication fork. This structure shows how RecG can recognize branched DNA structures and suggests a mechanism for fork reversal, an early event in recombination-dependent reinitiation of DNA replication.

Polyubiquitinated PCNA recruits the ZRANB3 translocase to maintain genomic integrity after replication stress.

Ciccia, Alberto Nimonkar, Amitabh V Hu, Yiduo Hajdu, Ildiko Achar, Yathish Jagadheesh Izhar, Lior Petit, Sarah A Adamson, Britt Yoon, John C Kowalczykowski, Stephen C ...

Published in Molecular cell

Completion of DNA replication after replication stress depends on PCNA, which undergoes monoubiquitination to stimulate direct bypass of DNA lesions by specialized DNA polymerases or is polyubiquitinated to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Here we report that the ZRANB3 translocase, ...

Direct visualization of RecBCD movement reveals cotranslocation of the RecD motor after chi recognition.

Naofumi Handa Bianco, Pr Baskin, Rj Stephen Charles Kowalczykowski

Published in Molecular Cell

In Escherichia coli, chi (5 -GCTGGTGG-3 ) is a recombination hotspot recognized by the RecBCD enzyme. Recognition of chi reduces both nuclease activity and translocation speed of RecBCD and activates RecA-loading ability. RecBCD has two motor subunits, RecB and RecD, which act simultaneously but independently. A longstanding hypothesis to explain t...

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