title: The HD-GYP domain of RpfG mediates a direct linkage between the Rpf quorum-sensing pathway and a subset of diguanylate cyclase proteins in the phytopathogen Xanthomonas axonopodis pv citri.
abstract: Bacteria use extracellular levels of small diffusible autoinducers to estimate local cell-density (quorum-sensing) and to regulate complex physiological processes. The quorum-sensing signal transduction pathway of Xanthomonas spp. phytopathogens has special features that distinguish it from that of other pathogens. This pathway consists of RpfF, necessary for the production of the unique autoinducer 'diffusible signalling factor' (DSF), and RpfC and RpfG, a two-component system necessary for the DSF-dependent production of extracellular pathogenicity factors and cellular dispersion. Yeast two-hybrid and direct in vitro assays were used to identify interactions involving the Rpf group of proteins. We show that RpfC, a protein consisting of N-terminal transmembrane, histidine kinase, response-regulator and C-terminal histidine phosphotransfer domains interacts with both RpfG, a protein consisting of an N-terminal response regulator domain and a C-terminal HD-GYP domain, and with RpfF. We also show that RpfC interacts with the only known homologue of 'conditioned medium factor', which is involved in quorum-sensing in Dictyostelium discoideum under conditions of nutritional stress. Furthermore, RpfCG is shown to interact with a second two-component system made up of NtrB and NtrC homologues. Finally we show that the recently characterized HD-GYP phosphodiesterase domain of RpfG interacts directly with diguanylate cyclase GGDEF domain-containing proteins coded by the Xanthomonas axonopodis pv. citri genome, which in other bacteria produce cyclic diGMP, an important second messenger involved in the regulation of complex bacterial processes including biofilm production, virulence and motility. These results demonstrate a direct physical linkage between quorum-sensing and cyclic diGMP signalling pathways in bacteria.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes, 748, CEP 05599-970 São Paulo, SP, Brazil.
authors.forename: Maxuel O
authors.initials: MO
authors.lastname: Andrade
authors.forename: Marcos Castanheira
authors.initials: MC
authors.lastname: Alegria
authors.forename: Cristiane R
authors.initials: CR
authors.lastname: Guzzo
authors.forename: Cassia
authors.initials: C
authors.lastname: Docena
authors.forename: Maria Claudia Pareda
authors.initials: MC
authors.lastname: Rosa
authors.forename: Carlos H I
authors.initials: CH
authors.lastname: Ramos
authors.forename: Chuck S
authors.initials: CS
authors.lastname: Farah
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D026481:Blotting, Far-Western / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008954:Models, Biological; D008969:Molecular Sequence Data; D019761:Phosphorus-Oxygen Lyases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D014974:Xanthomonas / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; C419552:RpfG protein, Xanthomonas campestris; C102500:rpfC protein, Xanthomonas; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase