Contributors: Biologie des Bactéries pathogènes à Gram-positif; Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS); Biologie Moléculaire du Gène chez les Extrêmophiles (BMGE); Institut Pasteur Paris (IP); Support was provided by Institut Pasteur Transversal Research Program No. 336; Agence Nationale de la Recherche, Grant ANR-08-ALIA-0011 NABAB; and Agence Nationale de la Recherche, Grant ANR-09-MIEN-0010 GRABIRON.; We are grateful to Dr. Stephen Leppla and Dr. Inka Sastalla for providing plasmid pTetONGFPopt. We thank Ons Ben Aïssa and Dr. Michel Débarbouillé for constructing walR and walJ transcriptional lacZ fusions, Dr. Gouzel Karimova for the kind gift of E. coli strain DHT1 and plasmids pKTop, pKT25 and pUT18c, Dr. Pascale Romby for RNA folding predictions and Adeline Mallet (Ultrapole, Institut Pasteur) for transmission electron micrographs.; ANR-08-ALIA-0011,NABAB,Stratégie Non AntiBiotique Anti-Bactéries pathogènes : exploration des capacités inhibitrices des écosystèmes naturels contre les contaminations à S. aureus en contexte laitier(2008); ANR-09-MIEN-0010,Grablron," Nouveaux systèmes d'acquisition du fer chez les pathogènes Gram+ extracellulaires "(2009)
نبذة مختصرة : International audience ; The WalKR two-component system, controlling cell wall metabolism, is highly conserved among Bacilli and essential for cell viability. In Staphylococcus aureus, walR and walK are followed by three genes of unknown function: walH, walI and walJ. Sequence analysis and transcript mapping revealed a unique genetic structure for this locus in S. aureus: the last gene of the locus, walJ, is transcribed independently, whereas transcription of the tetra-cis-tronic walRKHI operon occurred from two independent promoters located upstream from walR. Protein topology analysis and protein-protein interactions in E. coli as well as subcel-lular localization in S. aureus allowed us to show that WalH and WalI are membrane-bound proteins, which associate with WalK to form a complex at the cell division septum. While these interactions suggest that WalH and WalI play a role in activity of the WalKR regulatory pathway, deletion of walH and/or walI did not have a major effect on genes whose expression is strongly dependent on WalKR or on associated phenotypes. No effect of WalH or WalI was seen on tightly controlled WalKR regulon genes such as sle1 or saouhsc_00773, which encodes a CHAP-domain amidase. Of the genes encoding the two major S. aureus autolysins, AtlA and Sle1, only transcription of atlA was increased in the ΔwalH or ΔwalI mutants. Likewise, bacterial autolysis was not increased in the absence of WalH and/or WalI and biofilm formation was lowered rather than increased. Our results suggest that contrary to their major role as WalK inhibitors in B. subtilis, the WalH and WalI proteins have evolved a different function in S. aureus, where they are more accessory. A phylogenomic analysis shows a striking conservation of the 5 gene wal cluster along the evolutionary history of Bacilli, supporting the key importance of this signal transduction system, and indicating that the walH and walI genes were lost in the ancestor of Streptococcaceae, leading to their atypical 3 wal gene cluster, walRKJ.
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