Homologous recombination at the border: insertion-deletions and the trapping of foreign DNA in Streptococcus pneumoniae |
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Authors: | Prudhomme Marc Libante Virginie Claverys Jean-Pierre |
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Institution: | Laboratoire de Microbiologie et Génétique Moléculaire, Unité Mixte de Recherche 5100, Centre National de la Recherche Scientifique-Université Paul Sabatier, 118 Route de Narbonne, 31062 Toulouse Cedex, France. |
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Abstract: | Integration of foreign DNA was observed in the Gram-positive human pathogen Streptococcus pneumoniae (pneumococcus) after transformation with DNA from a recombinant Escherichia coli bacteriophage lamda carrying a pneumococcal insert. Segments of lamda DNA replaced chromosomal sequences adjacent to the region homologous with the pneumococcal insert, whence the name insertion-deletion. Here we report that a pneumococcal insert was absolutely required for insertion-deletion formation, but could be as short as 153 bp; that the sizes of foreign DNA insertions (289-2,474 bp) and concomitant chromosomal deletions (45-1,485 bp) were not obviously correlated; that novel joints clustered preferentially within segments of high GC content; and that the crossovers in 29 independent novel joints were located 1 bp from the border or within short (3-10 nt long) stretches of identity (microhomology) between resident and foreign DNA. The data are consistent with a model in which the insert serving as a homologous recombination anchor favors interaction and subsequent illegitimate recombination events at microhomologies between foreign and resident sequences. The potential of homology- directed illegitimate recombination for genome evolution was illustrated by the trapping of functional heterologous genes. |
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