Structural basis for the inhibition of RecBCD by Gam and its synergistic antibacterial effect with quinolones

Wilkinson, M. K.; Troman, Lucy; Ismah, Wan Ak Wan Nur; Chaban, Yuriy; Avison, Matthew B.; Dillingham, Mark S.; Wigley, Dale B.

doi:10.7554/elife.22963

Cited by 49 publications

(38 citation statements)

References 42 publications

(56 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…RecBCD has been validated as a target for antibiotic development (32). The differing modes of Chi interaction adopted by AddAB and RecBCD have important implications for drug discovery.…”

Section: Discussionmentioning

confidence: 99%

A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair

Cheng

Wilkinson

Chaban

et al. 2020

Nat Struct Mol Biol

Self Cite

View full text Add to dashboard Cite

The RecBCD complex plays key roles in phage DNA degradation, CRISPR array acquisition (adaptation) and host DNA repair. The switch between these roles is regulated by a DNA sequence called Chi. We report cryo-EM structures of the Escherichia coli RecBCD complex bound to several different DNA forks containing a Chi sequence, including one in which Chi is recognised and others in which it is not. The Chi-recognised structure shows conformational changes in regions of the protein that contact Chi and reveals a tortuous path taken by the DNA. Sequence specificity arises from interactions with both the RecC subunit and the sequence itself. These structures provide molecular details for how Chi is recognised and insights into the changes that occur in response to Chi binding that switch RecBCD from bacteriophage destruction and CRISPR spacer acquisition, to constructive host DNA repair.

show abstract

“…RecBCD has been validated as a target for antibiotic development (32). The differing modes of Chi interaction adopted by AddAB and RecBCD have important implications for drug discovery.…”

Section: Discussionmentioning

confidence: 99%

A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair

Cheng

Wilkinson

Chaban

et al. 2020

Nat Struct Mol Biol

Self Cite

View full text Add to dashboard Cite

show abstract

“…In most cases, annealing is thought to occur at a target site that is exposed as ssDNA on the lagging strand of a replication fork (7). A third protein, Gam, binds and inactivates the host RecBCD enzyme, to prevent it from degrading the phage genome (8). Interest in λ Exo and Redβ stems in part from their distant homologies with the mammalian recombination proteins Dna2 (9) and Rad52 (10–14), respectively.…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of the Redβ C-terminal domain in complex with λ Exonuclease reveals an unexpected homology with λ Orf and an interaction withEscherichia colisingle stranded DNA binding protein

Caldwell

Zakharova

Filsinger

et al. 2019

Nucleic Acids Research

View full text Add to dashboard Cite

Bacteriophage λ encodes a DNA recombination system that includes a 5′-3′ exonuclease (λ Exo) and a single strand annealing protein (Redβ). The two proteins form a complex that is thought to mediate loading of Redβ directly onto the single-stranded 3′-overhang generated by λ Exo. Here, we present a 2.3 Å crystal structure of the λ Exo trimer bound to three copies of the Redβ C-terminal domain (CTD). Mutation of residues at the hydrophobic core of the interface disrupts complex formation in vitro and impairs recombination in vivo . The Redβ CTD forms a three-helix bundle with unexpected structural homology to phage λ Orf, a protein that binds to E. coli single-stranded DNA binding protein (SSB) to function as a recombination mediator. Based on this relationship, we found that Redβ binds to full-length SSB, and to a peptide corresponding to its nine C-terminal residues, in an interaction that requires the CTD. These results suggest a dual role of the CTD, first in binding to λ Exo to facilitate loading of Redβ directly onto the initial single-stranded DNA (ssDNA) at a 3′-overhang, and second in binding to SSB to facilitate annealing of the overhang to SSB-coated ssDNA at the replication fork.

show abstract

“…Bet and Exo of SXT/R391 ICEs have been shown to promote hybrid ICE formation by RecA-independent homologous recombination between SXT and R391 integrated in a tandem fashion in the chromosome (21). Like IncC plasmids, SXT/R391 ICEs lack gam, which encodes an inhibitor of E. coli SbcCD and RecBCD exonuclease, but carry a ssb gene (46)(47)(48). Since SXT/R391 ICEs share homologous bet and exo genes (46), the role of this system was also investigated using R391.…”

Section: Ssb/bet/exo Allows Crispr-cas Evasion Of Ice R391 During Conmentioning

confidence: 99%

Faculty Opinions recommendation of IncC conjugative plasmids and SXT/R391 elements repair double-strand breaks caused by CRISPR-Cas during conjugation.

Bell

2020

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature

View full text Add to dashboard Cite

Structural basis for the inhibition of RecBCD by Gam and its synergistic antibacterial effect with quinolones

Cited by 49 publications

References 42 publications

A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair

A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair

Crystal structure of the Redβ C-terminal domain in complex with λ Exonuclease reveals an unexpected homology with λ Orf and an interaction withEscherichia colisingle stranded DNA binding protein

Faculty Opinions recommendation of IncC conjugative plasmids and SXT/R391 elements repair double-strand breaks caused by CRISPR-Cas during conjugation.

Contact Info

Product

Resources

About