Identification of the RecR Toprim Domain as the Binding Site for both RecF and RecO

Honda, Masayoshi; Inoue, Jin; Yoshimasu, Masatoshi; Ito, Yutaka; Shibata, Takehiko; Mikawa, Tsutomu

doi:10.1074/jbc.m512658200

Cited by 40 publications

(42 citation statements)

References 38 publications

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“…A DNA clamp function of RecR was suggested by structural studies of D. radiodurans RecR (45,46). A similar model of RecR-DNA interaction within the RecFR complex was proposed based on small angle x-ray scattering measurements of T. thermophilus proteins (52).…”

Section: Discussionmentioning

confidence: 99%

“…For T. thermophilus proteins, RecF and RecO binding sites on RecR overlap (46). Lack of structural similarity between RecF and RecO makes it difficult to predict which part of RecF binds RecR.…”

Section: Discussionmentioning

confidence: 99%

“…The E. coli RecR protein has never been reported to interact with DNA without RecF. The binding of D. radiodurans RecR to plasmid DNA in the presence of 40 mM Mg 2ϩ (45) and weak interactions of T. thermophilus RecR with DNA have been demonstrated (46). We tested whether a potential weak RecR affinity to DNA contributes to formation and stability of the RecF-DNA-RecR complex.…”

Section: Recr Stabilizes the Recf Dimerization And Dna Binding With Amentioning

confidence: 99%

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RecR-mediated Modulation of RecF Dimer Specificity for Single- and Double-stranded DNA

Makharashvili

Tian

Королева

et al. 2009

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Recr Stabilizes the Recf Dimerization And Dna Binding With Amentioning

confidence: 99%

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RecR-mediated Modulation of RecF Dimer Specificity for Single- and Double-stranded DNA

Makharashvili

Tian

Королева

et al. 2009

Journal of Biological Chemistry

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“…E. coli RecR protein binds to both RecF and RecO proteins in vitro (27,54,55) but not at the same time (47). The C-terminal TOPRIM domain of RecR has been implicated as the interaction site with both RecO and RecF, suggesting that these proteins may compete for RecR binding (56). The competition between RecF and RecO for RecR protein binding has been documented in vitro (47).…”

mentioning

confidence: 99%

SSB Protein Limits RecOR Binding onto Single-stranded DNA

Hobbs¹,

Sakai

Cox

2007

Journal of Biological Chemistry

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The RecO and RecR proteins form a complex that promotes the nucleation of RecA protein filaments onto SSB proteincoated single-stranded DNA (ssDNA). However, even when RecO and RecR proteins are provided at optimal concentrations, the loading of RecA protein is surprisingly slow, typically proceeding with a lag of 10 min or more. The rate-limiting step in RecOR-promoted RecA nucleation is the binding of RecOR protein to ssDNA, which is inhibited by SSB protein despite the documented interaction between RecO and SSB. Full activity of RecOR is seen only when RecOR is preincubated with ssDNA prior to the addition of SSB. The slow binding of RecOR to SSBcoated ssDNA involves the C terminus of SSB. When an SSB variant that lacks the C-terminal 8 amino acids is used, the capacity of RecOR to facilitate RecA loading onto the ssDNA is largely abolished. The results are used in an expanded model for RecOR action.

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“…E. coli RecR protein binds to both RecF and RecO proteins in vitro (31,[41][42][43]. There is an apparent competition between RecF and RecO for RecR binding that may involve an interaction of both RecF and RecO with the C-terminal TOPRIM domain of RecR (50,51). RecR increases the apparent affinity of both RecO and RecF for DNA (31,41,42).…”

mentioning

confidence: 99%

RecFOR and RecOR as Distinct RecA Loading Pathways

Sakai

Cox

2009

Journal of Biological Chemistry

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The molecular role of the RecF protein in loading RecA protein onto single-stranded DNA (ssDNA)-binding proteincoated ssDNA has been obscured by the facility with which the RecO and RecR proteins alone perform this function. We now show that RecFOR and RecOR define distinct RecA loading functions that operate optimally in different contexts. RecFOR, but not RecOR, is most effective when RecF(R) is bound near an ssDNA/double-stranded (dsDNA) junction. However, RecF(R) has no enhanced binding affinity for such a junction. RecO and RecR proteins are both required under all conditions in which the RecFOR pathway operates. The RecOR pathway is uniquely distinguished by a required interaction between RecO protein and the ssDNA binding protein C terminus. The RecOR pathway is more efficient for RecA loading onto ssDNA when no proximal dsDNA is available. A merger of new and published results leads to a new model for RecFOR function.Recombination reactions catalyzed by the RecA protein form an integral part of DNA metabolism in Escherichia coli. The major role for recombination in bacteria is the repair of stalled replication forks (1-4). E. coli strains lacking intact recombination systems exhibit sensitivity to DNA damaging agents, hypermutability, and impaired growth rates (5).RecA is found in all eubacteria with the exception of a few endosymbionts (Buchnera sp.) (6 -8). It functions as a helical nucleoprotein filament, formation of which requires ATP and Mg 2ϩ ion. When bound to single-stranded DNA (ssDNA), 2RecA hydrolyzes ATP. The active filament can promote the exchange of DNA strands between homologous DNA molecules in vitro. RecA filament formation occurs in at least two distinct phases (9). Filament nucleation occurs first and involves binding of a few (probably 4 -5 (10)) RecA protomers to DNA. Nucleation is rate-limiting and is followed by rapid filament extension in the 5Ј to 3Ј direction along the DNA (11-14). RecA filament disassembly requires ATP hydrolysis and also occurs in the 5Ј to 3Ј direction along ssDNA (14). Recombinase function is highly regulated in all cells, and the regulation occurs at many levels. The LexA repressor protein controls expression of the recA gene (15, 16). The 17 C-terminal amino acids of RecA function in autoregulation of most protein activities (17-19). Finally, E. coli possesses many additional proteins that function as recombination regulators, with RecA as their target (20 -22). Most of these proteins affect the kinetics of filament formation and disassembly. Proteins that facilitate the formation of the filaments of RecA-class recombinases are called recombination mediator proteins (21). Recombination mediator proteins are as ubiquitous as are the recombinases themselves (21,(23)(24)(25)(26)(27)(28). The E. coli RecF, RecO, and RecR proteins are perhaps the prototypical recombination mediator proteins (12, 21, 29 -32), part of a larger network of bacterial proteins that regulate almost every aspect of RecA function (22,30,33).The ssDNA-binding protein (SSB; 18.2 kDa) ex...

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Identification of the RecR Toprim Domain as the Binding Site for both RecF and RecO

Cited by 40 publications

References 38 publications

RecR-mediated Modulation of RecF Dimer Specificity for Single- and Double-stranded DNA

RecR-mediated Modulation of RecF Dimer Specificity for Single- and Double-stranded DNA

SSB Protein Limits RecOR Binding onto Single-stranded DNA

RecFOR and RecOR as Distinct RecA Loading Pathways

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