Bacterial RadA is a DnaB-type helicase interacting with RecA to promote bidirectional D-loop extension

Léa Marie, Chiara Rapisarda, Violette Morales, Mathieu Bergé, Thomas Perry, Anne-Lise Soulet, Clémence Gruget, Han Remaut, Rémi Fronzes () and Patrice Polard ()
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Léa Marie: Laboratoire de Microbiologie et Génétique Moléculaires, UMR5100, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS
Chiara Rapisarda: G5 Biologie structurale de la sécrétion bactérienne, UMR 3528, CNRS/Institut Pasteur, Institut Pasteur
Violette Morales: Laboratoire de Microbiologie et Génétique Moléculaires, UMR5100, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS
Mathieu Bergé: Laboratoire de Microbiologie et Génétique Moléculaires, UMR5100, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS
Thomas Perry: G5 Biologie structurale de la sécrétion bactérienne, UMR 3528, CNRS/Institut Pasteur, Institut Pasteur
Anne-Lise Soulet: Laboratoire de Microbiologie et Génétique Moléculaires, UMR5100, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS
Clémence Gruget: G5 Biologie structurale de la sécrétion bactérienne, UMR 3528, CNRS/Institut Pasteur, Institut Pasteur
Han Remaut: Structural and Molecular Microbiology, Structural Biology Research Center, VIB
Rémi Fronzes: G5 Biologie structurale de la sécrétion bactérienne, UMR 3528, CNRS/Institut Pasteur, Institut Pasteur
Patrice Polard: Laboratoire de Microbiologie et Génétique Moléculaires, UMR5100, Centre de Biologie Intégrative (CBI), Centre National de la Recherche Scientifique (CNRS), Université de Toulouse, UPS

Nature Communications, 2017, vol. 8, issue 1, 1-14

Abstract: Abstract Homologous recombination (HR) is a central process of genome biology driven by a conserved recombinase, which catalyses the pairing of single-stranded DNA (ssDNA) with double-stranded DNA to generate a D-loop intermediate. Bacterial RadA is a conserved HR effector acting with RecA recombinase to promote ssDNA integration. The mechanism of this RadA-mediated assistance to RecA is unknown. Here, we report functional and structural analyses of RadA from the human pathogen Streptococcus pneumoniae. RadA is found to facilitate RecA-driven ssDNA recombination over long genomic distances during natural transformation. RadA is revealed as a hexameric DnaB-type helicase, which interacts with RecA to promote orientated unwinding of branched DNA molecules mimicking D-loop boundaries. These findings support a model of DNA branch migration in HR, relying on RecA-mediated loading of RadA hexamers on each strand of the recipient dsDNA in the D-loop, from which they migrate divergently to facilitate incorporation of invading ssDNA.

Date: 2017
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DOI: 10.1038/ncomms15638

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