Resolution of Holliday junctions in Escherichia coli: identification of the ruvC gene product as a 19-kilodalton protein

Sharples, Gary J.; Lloyd, Robert G.

doi:10.1128/jb.173.23.7711-7715.1991

Cited by 80 publications

(38 citation statements)

References 31 publications

(25 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This clone does not fully restore UV resistance to ruv mutants, possibly due to a slight negative effect on cell survival after UV exposure. Other plasmids used were pGS762, pGS711, and pPVA101 (Sharples et al 1990;Sharples and Lloyd 1991).…”

Section: Methodsmentioning

confidence: 99%

The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis

et al. 2005

View full text Add to dashboard Cite

In models of Escherichia coli recombination and DNA repair, the RuvABC complex directs the branch migration and resolution of Holliday junction DNA. To probe the validity of the E. coli paradigm, we examined the impact of mutations in DruvAB and DrecU (a ruvC functional analog) on DNA repair. Under standard transformation conditions we failed to construct DruvAB DrecG, DrecU DruvAB, DrecU DrecG, or DrecU DrecJ strains. However, DruvAB could be combined with addAB (recBCD), recF, recH, DrecS, DrecQ, and DrecJ mutations. The DruvAB and DrecU mutations rendered cells extremely sensitive to DNA-damaging agents, although less sensitive than a DrecA strain. When damaged cells were analyzed, we found that RecU was recruited to defined double-stranded DNA breaks (DSBs) and colocalized with RecN. RecU localized to these centers at a later time point during DSB repair, and formation was dependent on RuvAB. In addition, expression of RecU in an E. coli ruvC mutant restored full resistance to UV light only when the ruvAB genes were present. The results demonstrate that, as with E. coli RuvABC, RuvAB targets RecU to recombination intermediates and that all three proteins are required for repair of DSBs arising from lesions in chromosomal DNA.

show abstract

Section: Methodsmentioning

confidence: 99%

The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis

et al. 2005

View full text Add to dashboard Cite

show abstract

“…The product of the ruvC gene, which is not regulated by SOS system (Sharples et al, 1990;Sharples and Lloyd, 1991;Takahagi et al, 1991), resolves Holliday junctions to give mature recombinant DNA molecules by introducing nicks at the crossover junction in the strands with the same polarity (Dunderdale et al, 1991;Iwasaki et al, 1991;Sharples and Lloyd, 1991). The role of RecG in recombination is less well understood.…”

Section: Introductionmentioning

confidence: 99%

Roles of the recG gene product of Escherichia coli in recombination repair: Effects of the .DELTA.recG mutation on cell division and chromosome partition.

1997

View full text Add to dashboard Cite

The products of the recG and ruvAB genes of Escherichia coli are both thought to promote branch migration of Holliday recombination intermediates by their junction specific helicase activities in homologous recombination and recombination repair. To investigate the in vivo role of the recG gene, we examined the effects of a recG null mutation on cell division and chromosome partition. After UV irradiation at a low dose (5J/ m 2 ), ∆recG mutant formed filamentous cells with unpartitioned chromosomes. A mutation in the sfiA gene, which encodes an SOS-inducible inhibitor of septum formation, partially suppressed filamentation of recG mutant cells, but did not prevent the formation of anucleate cells. The sensitivity to UV light and the cytological phenotypes after UV irradiation of a recA recG double mutant were similar to a recA single mutant, consistent with the role of recG, which is assigned to a later stage in recombination repair than recA. The recG ruvAB and recG ruvC double mutants were more sensitive to UV, almost as sensitive as the recA mutant and showed more extreme phenotypes concerning filamentation and chromosome nondisjunction, both after UV irradiation and without UV irradiation than either recG or ruv single mutants. The recG polA12 (Ts) mutant, which is temperature sensitive in growth, formed filamentous cells with centrally located chromosome aggregates when grown at nonpermissive temperature similar to the UV irradiated recG mutant. These results support the notion that RecG is involved in processing Holliday intermediates in recombination repair in vivo. We suggest that the defect in the processing in the recG mutant results in accumulation of nonpartitioned chromosomes, which are linked by Holliday junctions.

show abstract

“…and ruvB genes constitute a LexA-regulated operon [Shurvinton and Lloyd 1982;Benson et al 1988;Shinagawa et al 1988), whereas the ruvC gene located near the ruvA-ruvB operon is not regulated by the SOS system (Sharpies et al 1990;Sharpies and Lloyd 1991;Takahagi et al 1991). Like the recA gene, the ruvA and ruvB genes apparently play multiple roles in recombination, repair, and mutagenesis.…”

mentioning

confidence: 99%

Escherichia coli RuvA and RuvB proteins specifically interact with Holliday junctions and promote branch migration.

Iwasaki¹,

Takahagi²,

Nakata³

et al. 1992

Genes Dev.

174

131

View full text Add to dashboard Cite

The Escherichia coli ruvA and ruvB genes are involved in DNA repair and in the late step of homologous genetic recombination. We have demonstrated previously that the RuvA-RuvB protein complex in the presence of ATP promotes reabsorption of cruciform structures extruded from a supercoiled plasmid with an inverted repeat sequence. Because the cruciform structure is topologically analogous to the Holliday structure, we have proposed that the role of the RuvA and RuvB proteins in recombination is to promote a strand exchange reaction at the Holliday junction. Here, we studied the specific interaction of the RuvA-RuvB complex with the Holliday structure using synthetic analogs prepared by annealing four oligonucleotides. The affinities of the RuvA protein for synthetic Holliday junctions are much higher (>20-fold) than for duplex DNA, and the affinities of the RuvA protein for the junctions are further enhanced (>4-fold) by the interaction with the RuvB protein. The RuvA-RuvB protein complex in the presence of ATP promotes dissociation of the synthetic Holliday junction with homology in the central core into two halves by catalyzing branch migration to the DNA ends, but it does not affect the structure of the synthetic Holliday junction without the homology. The separation of the synthetic Holliday junction is a result of the activity of the RuvA-RuvB complex that promotes strand exchange and DNA unwinding. Furthermore, RuvA and RuvB promote the strand exchange reaction at the Holliday junctions made by RecA. These results provide further evidence that the RuvA-RuvB complex recognizes the Holliday junction and promotes branch migration in homologous recombination.

show abstract

Resolution of Holliday junctions in Escherichia coli: identification of the ruvC gene product as a 19-kilodalton protein

Cited by 80 publications

References 31 publications

The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis

The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis

Roles of the recG gene product of Escherichia coli in recombination repair: Effects of the .DELTA.recG mutation on cell division and chromosome partition.

Escherichia coli RuvA and RuvB proteins specifically interact with Holliday junctions and promote branch migration.

Contact Info

Product

Resources

About