Structure-function analysis of pneumococcal DprA protein reveals that dimerization is crucial for loading RecA recombinase onto DNA during transformation.

Abstract:

:Transformation promotes genome plasticity in bacteria via RecA-driven homologous recombination. In the gram-positive human pathogen Streptococcus pneumoniae, the transformasome a multiprotein complex, internalizes, protects, and processes transforming DNA to generate chromosomal recombinants. Double-stranded DNA is internalized as single strands, onto which the transformation-dedicated DNA processing protein A (DprA) ensures the loading of RecA to form presynaptic filaments. We report that the structure of DprA consists of the association of a sterile alpha motif domain and a Rossmann fold and that DprA forms tail-to-tail dimers. The isolation of DprA self-interaction mutants revealed that dimerization is crucial for the formation of nucleocomplexes in vitro and for genetic transformation. Residues important for DprA-RecA interaction also were identified and mutated, establishing this interaction as equally important for transformation. Positioning of key interaction residues on the DprA structure revealed an overlap of DprA-DprA and DprA-RecA interaction surfaces. We propose a model in which RecA interaction promotes rearrangement or disruption of the DprA dimer, enabling the subsequent nucleation of RecA and its polymerization onto ssDNA.

authors

Quevillon-Cheruel S,Campo N,Mirouze N,Mortier-Barrière I,Brooks MA,Boudes M,Durand D,Soulet AL,Lisboa J,Noirot P,Martin B,van Tilbeurgh H,Noirot-Gros MF,Claverys JP,Polard P

doi

10.1073/pnas.1205638109

subject

Has Abstract

pub_date

2012-09-11 00:00:00

pages

E2466-75

issue

37

eissn

0027-8424

issn

1091-6490

pii

1205638109

journal_volume

109

pub_type

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