Proteomic analysis of quorum sensing in Rhizobium leguminosarum biovar viciae UPM791

Abstract

Production of quorum-sensing signal molecules of the acyl-homoserine lactone (AHL) type by Rhizobium leguminosarum bv. viciae UPM791 is dependent on its plasmid content. Curing of two of its four native plasmids, pUPM791d and pSym, resulted in loss of production of the largest (C(14)) and the three smaller (C(6)-C(8)) AHLs, respectively. Introduction of a lactonase-containing plasmid resulted in AHL signal degradation and quorum quenching. The quorum-dependent proteome was studied in these strains by DIGE. Quorum quenching affected a small (1.7%) fraction of the detected spots in the wild-type and a smaller (0.6%) fraction in the pSym-cured strain. Unexpectedly, quorum quenching affected up to 3.3% of the detected spots in the pUPM791d-cured strain, suggesting that C(14)-AHL normally interferes with the quorum response mediated by other AHLs. This, together with the observation that ca. 50% of the quorum-regulated proteins in strain UPM791 showed AHL-mediated repression, suggests that an important part of their functionality can be exerted through repression, although AHLs are usually considered as gene expression inducers. The three main quorum-induced polypeptides were identified by MALDI-MS as charge isoforms of the rhizospheric RhiA protein. Another major quorum-induced polypeptide was only present in the pUPM791d-cured strain and could not be identified.