Biochemical basis of hyper‐recombinogenic activity of <i>Pseudomonas aeruginosa</i> RecA protein in <i>Escherichia coli</i> cells

Namsaraev, Eugene A.; Baitin, Dmitry; Bakhlanova, Irina V.; Alexseyev, Andrey A.; Ogawa, Hideyuki; Lanzov, Vladislav A.

doi:10.1046/j.1365-2958.1998.00718.x

Cited by 23 publications

(32 citation statements)

References 30 publications

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“…In addition to TLS induction, are there other functional differences between RecA C and RecAP? They likely have largely overlapping functions, but RecAC and RecAP may be optimized for different activities in for example homologous recombination, repair of stalled replication forks or RecA-mediated excision repair (Ishimori et al, 1996;Namsaraev et al, 1998;Bichara et al, 2007;Cox, 2007a,b). Furthermore, RecAC and RecAP may have yet unidentified different coprotease targets.…”

Section: Discussionmentioning

confidence: 99%

Mutagenic lesion bypass and two functionally different RecA proteins in Deinococcus deserti

Dulermo

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Blanchard

et al. 2009

Molecular Microbiology

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SummaryRecA is essential for extreme radiation tolerance in Deinococcus radiodurans. Interestingly, Sahara bacterium Deinococcus deserti has three recA genes (recAC, recAP1, recAP3) that code for two different RecA proteins (RecAC, RecAP). Moreover, and in contrast to other sequenced Deinococcus species, D. deserti possesses homologues of translesion synthesis (TLS) DNA polymerases, including ImuY and DnaE2. Together with a lexA homologue, imuY and dnaE2 form a gene cluster similar to a widespread RecA/LexA-controlled mutagenesis cassette. After having developed genetic tools, we have constructed mutant strains to characterize these recA and TLS polymerase genes in D. deserti. Both RecAC and RecAP are functional and allow D. deserti to survive, and thus repair massive DNA damage, after exposure to high doses of radiation. D. deserti is mutable by UV, which requires ImuY, DnaE2 and RecAC, but not RecAP. RecAC, but not RecAP, facilitates induced expression of imuY and dnaE2 following UV exposure. We propose that the extra recAP1 and recAP3 genes may provide higher levels of RecA protein for efficient error-free repair of DNA damage, without further increasing error-prone lesion bypass by ImuY and DnaE2, whereas limited TLS may contribute to adaptation to harsh conditions by generating genetic variability.

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Section: Discussionmentioning

confidence: 99%

Mutagenic lesion bypass and two functionally different RecA proteins in Deinococcus deserti

Dulermo

Fochesato

Blanchard

et al. 2009

Molecular Microbiology

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“…Recombination-deficient strains JC7623-F, JC7623-O, and JC7623-R were constructed by P1 transduction to transfer recO1504::Tn5 (36), recR252::Tn10-9 (33), and recF349(del) (47) mutant genes, respectively, into strain JC7623. Plasmids pRecAEc (pUC19-recA1.1) and pRecAPa (pUC19-PA189.1) contain recA Ec and recA Pa genes, respectively, together with their native operator-promoter regions as described previously (37). Plasmid P200 FЈ-lac was used to standardize conjugation abilities of recipient strains.…”

Section: Methodsmentioning

confidence: 99%

“…The RecA Ec and RecA Pa proteins were purified as described previously (37 Conjugation. Conjugation was carried out essentially as described previously (25).…”

Section: Methodsmentioning

confidence: 99%

“…This enhancement of recombination is SOS independent (5). In E. coli, expression of RecA Pa increases FRE by six-to eightfold (37). RecA Pa displaces both the E. coli SSB and P. aeruginosa SSB from ssDNA more efficiently than RecA Ec does (4).…”

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confidence: 99%

“…It possesses a greater affinity for ssDNA than RecA Ec does (4). It promotes faster joint molecule formation in DNA strand exchange reactions but is less effective in generating the final products of DNA strand exchange (37). The propensity of RecA Pa to initiate but not complete strand exchange is a characteristic of recombinase nucleoprotein filaments with enhanced DNA pairing and/or duplex DNA binding activities (21,27).…”

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confidence: 99%

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Distinguishing Characteristics of Hyperrecombinogenic RecA Protein from Pseudomonas aeruginosa Acting in Escherichia coli

et al. 2006

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In Escherichia coli, a relatively low frequency of recombination exchanges (FRE) is predetermined by the activity of RecA protein, as modulated by a complex regulatory program involving both autoregulation and other factors. The RecA protein of Pseudomonas aeruginosa (RecA Pa ) exhibits a more robust recombinase activity than its E. coli counterpart (RecA Ec ). Low-level expression of RecA Pa in E. coli cells results in hyperrecombination (an increase of FRE) even in the presence of RecA Ec . This genetic effect is supported by the biochemical finding that the RecA Pa protein is more efficient in filament formation than RecA K72R, a mutant protein with RecA Ec -like DNA-binding ability. Expression of RecA Pa also partially suppresses the effects of recF, recO, and recR mutations. In concordance with the latter, RecA Pa filaments initiate recombination equally from both the 5 and 3 ends. Besides, these filaments exhibit more resistance to disassembly from the 5 ends that makes the ends potentially appropriate for initiation of strand exchange. These comparative genetic and biochemical characteristics reveal that multiple levels are used by bacteria for a programmed regulation of their recombination activities.

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Conformational flexibility of RecA protein filament: Transitions between compressed and stretched states

et al. 2006

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RecA protein is a central enzyme in homologous DNA recombination, repair and other forms of DNA metabolism in bacteria. It functions as a flexible helix-shaped filament bound on stretched single-stranded or double-stranded DNA in the presence of ATP. In this work, we present an atomic level model for conformational transitions of the RecA filament. The model describes small movements of the RecA N-terminal domain due to coordinated rotation of main chain dihedral angles of two amino acid residues (Psi/Lys23 and Phi/Gly24), while maintaining unchanged the RecA intersubunit interface. The model is able to reproduce a wide range of observed helix pitches in transitions between compressed and stretched conformations of the RecA filament. Predictions of the model are in agreement with Small Angle Neutron Scattering (SANS) measurements of the filament helix pitch in RecA::ADP-AlF(4) complex at various salt concentrations.

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Biochemical basis of hyper‐recombinogenic activity of Pseudomonas aeruginosa RecA protein in Escherichia coli cells

Cited by 23 publications

References 30 publications

Mutagenic lesion bypass and two functionally different RecA proteins in Deinococcus deserti

Mutagenic lesion bypass and two functionally different RecA proteins in Deinococcus deserti

Distinguishing Characteristics of Hyperrecombinogenic RecA Protein from Pseudomonas aeruginosa Acting in Escherichia coli

Conformational flexibility of RecA protein filament: Transitions between compressed and stretched states

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