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

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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
    Present address: Institut Européen de Chimie et Biologie, UMR 5234 Microbiologie fondamentale et pathogénicité CNRS/université de Bordeaux, 2 rue Robert Escarpit, 33607 Pessac, France)

  • 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
    Present address: Institut Européen de Chimie et Biologie, UMR 5234 Microbiologie fondamentale et pathogénicité CNRS/université de Bordeaux, 2 rue Robert Escarpit, 33607 Pessac, France)

  • 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
    Present address: Institut Européen de Chimie et Biologie, UMR 5234 Microbiologie fondamentale et pathogénicité CNRS/université de Bordeaux, 2 rue Robert Escarpit, 33607 Pessac, France)

  • 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)

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.

Suggested Citation

  • Léa Marie & Chiara Rapisarda & Violette Morales & Mathieu Bergé & Thomas Perry & Anne-Lise Soulet & Clémence Gruget & Han Remaut & Rémi Fronzes & Patrice Polard, 2017. "Bacterial RadA is a DnaB-type helicase interacting with RecA to promote bidirectional D-loop extension," Nature Communications, Nature, vol. 8(1), pages 1-14, August.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms15638
    DOI: 10.1038/ncomms15638
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    Cited by:

    1. Prashant P. Damke & Louisa Celma & Sumedha M. Kondekar & Anne Marie Di Guilmi & Stéphanie Marsin & Jordane Dépagne & Xavier Veaute & Pierre Legrand & Hélène Walbott & Julien Vercruyssen & Raphaël Guér, 2022. "ComFC mediates transport and handling of single-stranded DNA during natural transformation," Nature Communications, Nature, vol. 13(1), pages 1-13, December.

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