Bacillus subtilis DprA Recruits RecA onto Single-stranded DNA and Mediates Annealing of Complementary Strands Coated by SsbB and SsbA*

Background: Different mediators assist RecA to catalyze genetic recombination. Results: DprA facilitates the displacement of both SSBs (SsbB and SsbA), increases RecA nucleation onto SSB-coated ssDNA, and mediates DNA strand annealing. Conclusion: DprA facilitates RecA-mediated strand exchange and anneals complementary strands coated by an SSB protein. Significance: RecA-dependent chromosomal transformation and RecA-independent plasmid transformation require the competence-induced DprA mediator. Naturally transformable bacteria recombine internalized ssDNA with a homologous resident duplex (chromosomal transformation) or complementary internalized ssDNAs (plasmid or viral transformation). Bacillus subtilis competence-induced DprA, RecA, SsbB, and SsbA proteins are involved in the early processing of the internalized ssDNA, with DprA physically interacting with RecA. SsbB and SsbA bind and melt secondary structures in ssDNA but limit RecA loading onto ssDNA. DprA binds to ssDNA and facilitates partial dislodging of both single-stranded binding (SSB) proteins from ssDNA. In the absence of homologous duplex DNA, DprA does not significantly increase RecA nucleation onto protein-free ssDNA. DprA facilitates RecA nucleation and filament extension onto SsbB-coated or SsbB plus SsbA-coated ssDNA. DprA facilitates RecA-mediated DNA strand exchange in the presence of both SSB proteins. DprA, which plays a crucial role in plasmid transformation, anneals complementary strands preferentially coated by SsbB to form duplex circular plasmid molecules. Our results provide a mechanistic framework for conceptualizing the coordinated events modulated by SsbB in concert with SsbA and DprA that are crucial for RecA-dependent chromosomal transformation and RecA-independent plasmid transformation.

• Publication year

  2013

• Venue

  Journal of Biological Chemistry

• Publication date

  2013-06-18

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1074/jbc.M113.478347PMID 23779106PMCID PMC3829333
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• DprA mediates annealing of complementary strands preferentially coated by SsbB to form circular plasmid duplexes.

  Confidence 0.97

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• DprA facilitates RecA-mediated DNA strand exchange when ssDNA is coated by both SsbB and SsbA.

  Confidence 0.96

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• DprA partially dislodges both SsbB and SsbA from single-stranded DNA and promotes RecA nucleation and filament extension on SSB-coated ssDNA.

  Confidence 0.98

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review

• dna strand annealing

  molecular process

  The pairing of complementary single-stranded DNA molecules into a duplex.

  Aliases: annealing

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• dna strand exchange

  molecular process

  The RecA-driven recombination reaction in which a single strand pairs and exchanges with homologous duplex DNA.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• dpra

  protein

  A competence-induced Bacillus subtilis mediator protein that binds ssDNA and interacts with RecA.

  Aliases: competence-induced DprA

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• plasmid transformation

  biological process

  A RecA-independent transformation route in which complementary internalized ssDNAs anneal to form circular plasmid molecules.

  Aliases: plasmid or viral transformation

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• reca

  protein

  The bacterial recombinase protein that catalyzes homologous recombination and strand exchange on DNA substrates.

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• single-stranded dna

  nucleic acid

  The internalized DNA substrate in single-stranded form that is coated by SSB proteins in the competence pathway.

  Aliases: ssDNA

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• ssba

  protein

  A Bacillus subtilis single-stranded DNA-binding protein that binds exposed ssDNA and is present alongside SsbB.

  Aliases: single-stranded binding protein A

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review
• ssbb

  protein

  A Bacillus subtilis single-stranded DNA-binding protein associated with competence and ssDNA coating.

  Aliases: single-stranded binding protein B

  박진우 (dztg5apj7m) extractionB (s683577b42) reviewAnonymous (12632b8b5f) review

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