Transformation of Streptococcus pneumoniae relies on DprA- and RecA-dependent protection of incoming DNA single strands.
Seventy-five years after the discovery of transformation with Streptococcus pneumoniae, it is remarkable how little we know of the proteins that interact with incoming single strands in the early processing of transforming DNA. In this work, we used as donor DNA in transformation a radioactively labelled homologous fragment to examine the fate of the single-stranded (ssDNA) products of uptake in cells mutant for DprA or RecA, two proteins essential for transformation. Fifteen minutes after uptake, the labelling of specific chromosomal restriction fragments that demonstrated homologous integration in the wild type was not detected in dprA or recA cells, indicating that in the mutants incoming ssDNA could not be processed into recombinants. Investigation of the fate of donor label 1 min after uptake revealed that incoming ssDNA was immediately degraded in the absence of DprA or RecA. Our results demonstrate that incoming ssDNA requires active protection prior to the RecA-driven search for homology and that both DprA and RecA are needed for this protection.
Bergé
Mathieu
M
Laboratoire de Microbiologie et Génétique Moléculaires, UMR 5100 CNRS-Université Paul Sabatier, 118 route de Narbonne, 31062 Toulouse Cedex, France.
Mortier-Barrière
Isabelle
I
Martin
Bernard
B
Claverys
Jean-Pierre
JP
D001426:Bacterial Proteins / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D011938:Rec A Recombinases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011995:Recombination, Genetic / Q000502:physiology; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial / Q000502:physiology*; D014170:Transformation, Genetic / Q000502:physiology*
D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; C093815:DprA protein, bacteria; D008565:Membrane Proteins; D011938:Rec A Recombinases