Proceedings of the National Academy of Sciences - PNAS, 2012-09, Vol.109 (37), p.E2466-E2475
2012
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- Autor(en) / Beteiligte
- Titel
- Structure–function analysis of pneumococcal DprA protein reveals that dimerization is crucial for loading RecA recombinase onto DNA during transformation
- Ist Teil von
- Proceedings of the National Academy of Sciences - PNAS, 2012-09, Vol.109 (37), p.E2466-E2475
- Ort / Verlag
- United States: National Acad Sciences
- Erscheinungsjahr
- 2012
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0027-8424eISSN: 1091-6490DOI: 10.1073/pnas.1205638109Titel-ID: cdi_hal_primary_oai_HAL_hal_01004530v1
- Format
- –
- Schlagworte
- Agricultural sciences, Bacterial proteins, Bacterial Proteins - chemistry, Bacterial Proteins - metabolism, Biological Sciences, Blotting, Western, Crystallization, Deoxyribonucleic acid, Dimerization, DNA, DNA - metabolism, DNA Primers - genetics, Genomes, Gram-positive bacteria, Life Sciences, Membrane Proteins - chemistry, Membrane Proteins - metabolism, Models, Molecular, Mutagenesis, Site-Directed, Mutation, PNAS Plus, Protein Conformation, Rec A Recombinases - metabolism, Streptococcus infections, Streptococcus pneumoniae - metabolism, Transformation, Bacterial - genetics, Transformation, Bacterial - physiology, Two-Hybrid System Techniques