Role of the Single-Stranded DNA–Binding Protein SsbB in Pneumococcal Transformation Maintenance of a Reservoir for Genetic Plasticity 英文参考文献.docVIP

RoleoftheSingle-StrandedDNA–BindingProteinSsbB inPneumococcalTransformation:Maintenanceofa ReservoirforGeneticPlasticity LaetitiaAttaiech1,2.,AudreyOlivier1,2.,IsabelleMortier-Barrie`re1,2.,Anne-LiseSoulet1,2 ,Chantal Granadel1,2,BernardMartin1,2,PatricePolard1,2,Jean-PierreClaverys1,2 * 1Centre National de la Recherche Scientifique, LMGM-UMR5100, Toulouse, France, 2Universite′ de Toulouse, UPS, Laboratoire de Microbiologie et Ge′ne′tique Mole′culaires,Toulouse,France Abstract Bacteria encode a single-stranded DNA (ssDNA) binding protein (SSB) crucial for genome maintenance. In Bacillus subtilis and Streptococcus pneumoniae,analternative SSB, SsbB, is expressed uniquely during competence for genetic transformation, but its precise role has been disappointingly obscure. Here, we report our investigations involving comparison of anull mutant (ssbB2)and aC-ter truncation (ssbBD7)of SsbB of S. pneumoniae,the latter constructed because SSBs’ acidic tail has emerged as a key site for interactions with partner proteins. We provide evidence that SsbB directly protects internalized ssDNA. We show that SsbB is highly abundant, potentially allowing the binding of ,1.15 Mb ssDNA (half a genome equivalent); that it participates in the processing of ssDNA into recombinants; and that, at high DNA concentration, it is of crucial importance for chromosomal transformation whilst antagonizing plasmid transformation. While thelatter observation explains along-standing observation thatplasmid transformation is very inefficient in S. pneumoniae (compared to chromosomal transformation), the former supports our previous suggestion that SsbB creates a reservoir of ssDNA, allowing successive recombination cycles. SsbBD7 fulfils the reservoir function, suggesting that SsbB C-ter is not necessary for processing protein(s) to access stored ssDNA. We propose that the evolutionary raison d’e?tre of SsbB and its abundance is maintenance of this reservoir, which contributes tothegenetic plasti