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To understand how the Rhizobium leguminosarum rail-raiR quorum-sensing system is regulated, we identified mutants with decreased levels of RaiI-made N-acyl homoserine lactones (AHLs). A LuxR-type regulator, ExpR, is required for raiR expression, and RaiR is required to induce rail. Since raiR (and rail) expression is also reduced in cinI and cinR quorum-sensing mutants, we thought CinI-made AHLs may activate ExpR to induce raiR. However, added CinI-made AHLs did not induce raiR expression in a cinI mutant. The reduced raiR expression in cinI and cinR mutants was due to lack of expression of cinS immediately downstream of cinI. cinS encodes a 67-residue protein, translationally coupled to CinI, and cinS acts downstream of expR for raiR induction. Cloned cinS in R. leguminosarum caused an unusual collapse of colony structure, and this was delayed by mutation of expR. The phenotype looked like a loss of exopolysaccharide (EPS) integrity; mutations in cinI, cinR, cinS, and expR all reduced expression of plyB, encoding an EPS glycanase, and mutation of plyB abolished the effect of cloned cinS on colony morphology. We conclude that CinS and ExpR act to increase PlyB levels, thereby influencing the bacterial surface. CinS is conserved in other rhizobia, including Rhizobium etli; the previously observed effect of cinI and cinR mutations decreasing swarming in that strain is primarily due to a lack of CinS rather than a lack of CinI-made AHL. We conclude that CinS mediates quorum-sensing regulation because it is coregulated with an AHL synthase and demonstrate that its regulatory effects can occur in the absence of AHLs. Copyright © 2009, American Society for Microbiology.


Documento: Artículo
Título:The cin and rai quorum-sensing regulatory systems in Rhizobium leguminosarum are coordinated by ExpR and CinS, a small regulatory protein coexpressed with CinI
Autor:Edwards, A.; Frederix, M.; Wisniewski-Dyé, F.; Jones, J.; Zorreguieta, A.; Allan Downie, J.
Filiación:John Innes Centre, Colney Norwich NR4 7UH, United Kingdom
Université Lyon 1, CNRS UMR 5557 Ecologie Microbienne, 69622 Villeurbanne, France
Fundación Instituto Leloir, IIBBA-CONICET, FCEyN, University of Buenos Aires, Patricias Argentinas 435, Buenos Aires, Argentina
John Innes Centre, Norwich Research Park, Colney Norwich NR4 7 UH, United Kingdom
Palabras clave:exopolysaccharide; glucan synthase; n acylhomoserine lactone; protein; regulator protein; regulatory protein Cin; regulatory protein CinI; regulatory protein CinS; regulatory protein ExpR; regulatory protein plyB; regulatory protein Rai; regulatory protein RaiR; unclassified drug; bacterial protein; gamma butyrolactone derivative; transactivator protein; animal experiment; article; bacterial spore; bacterial strain; cloning; colony formation; controlled study; cross coupling reaction; downstream processing; enzyme activity; gene identification; gene mutation; micromorphology; mutant; nonhuman; nucleotide sequence; phenotype; priority journal; protein expression; protein induction; protein structure; quorum sensing; regulatory mechanism; residue analysis; Rhizobiaceae; Rhizobium etli; Rhizobium leguminosarum; biological model; biosynthesis; gene expression regulation; metabolism; physiology; reporter gene; Bacteria (microorganisms); Rhizobium etli; Rhizobium leguminosarum; Acyl-Butyrolactones; Bacterial Proteins; Gene Expression Regulation, Bacterial; Genes, Reporter; Models, Biological; Quorum Sensing; Rhizobium leguminosarum; Trans-Activators
Página de inicio:3059
Página de fin:3067
Título revista:Journal of Bacteriology
Título revista abreviado:J. Bacteriol.
CAS:glucan synthase, 55126-98-2, 74191-29-0; protein, 67254-75-5; Acyl-Butyrolactones; Bacterial Proteins; Trans-Activators


  • Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., Lipman, D.J., Gapped BLAST and PSI-BLAST: A new generation of protein database search programs (1997) Nucleic Acids Res, 25, pp. 3389-3402
  • Bagdasarian, M., Lurz, R., Ruckert, B., Franklin, F.C.H., Bagdasar- ian, M.M., Frey, J., Timmis, K.N., Specific-purpose plasmid cloning vectors: 2 broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas (1981) Gene, 16, pp. 237-247
  • Becker, A., Puhler, A., Production of exopolysaccharides (1998) The Rhizo- biaceae, pp. 97-118. , H. P. Spaink, A. Kondorosi, and P. J. J. Hooykaas ed, Kluwer Academic, Dordrecht, The Netherlands
  • Beringer, J.E., R factor transfer in Rhizobium leguminosarum (1974) J. Gen. Microbiol, 84, pp. 188-198
  • Braeken, K., Daniels, R., Vos, K., Fauvart, M., Bachaspatimayum, D., Vanderleyden, J., Michiels, J., Genetic determinants of swarming in Rhizobium etli (2008) Microb. Ecol, 55, pp. 54-64
  • Buchanan-Wollaston, A.V., Generalised transduction in Rhizobium leguminosarum (1979) J. Gen. Microbiol, 112, pp. 135-142
  • Cha,C.,P.Gao,Y.C.Chen,P.D.Shaw,andS.K.Farrand.1998.Production of acyl-homoserine lactone quorum-sensing signals by gram-negative plant- associated bacteria. Mol. Plant-Microbe Interact. 11:1119-1129; Cubo, M.T., Economou, A., Murphy, G., Johnston, A.W., Downie, J.A., Molecular characterization and regulation of the rhizosphere-ex- pressed genes rhiABCR that can influence nodulation by Rhizobium legu- minosarum biovar viciae (1992) J. Bacteriol, 174, pp. 4026-4035
  • Daniels, R., De Vos, D.E., Desair, J., Raedschelders, G., Luyten, E., Rose- meyer, V., Verreth, C., Michiels, J., The cin quorum sensing locus of Rhizobium etli CNPAF512 affects growth and symbiotic nitrogen fixation (2002) J. Biol. Chem, 277, pp. 462-468
  • Daniels, R., Reynaert, S., Hoekstra, H., Verreth, C., Janssens, J., Braeken, K., Fauvart, M., Michiels, J., Quorum signal molecules as biosurfactants affecting swarming in Rhizobium etli (2006) Proc. Natl. Acad. Sci. USA, 103, pp. 14965-14970
  • Daniels, R., Vanderleyden, J., Michiels, J., Quorum sensing and swarming migration in bacteria (2004) FEMS Microbiol. Rev, 28, pp. 261-289
  • Danino, V.E., Wilkinson, A., Edwards, A., Downie, J.A., Recipient- induced transfer of the symbiotic plasmid pRL1JI in Rhizobium legumino- sarum bv. viciae is regulated by a quorum-sensing relay (2003) Mol. Microbiol, 50, pp. 511-525
  • Downie, J.A., Gonzalez, J.E., Cell-to-cell communication in rhizobia: Quorum sensing and plant signalling (2008) Chemical communication among bacteria, pp. 213-232. , S. C. Winans and B. L. Bassler ed, ASM Press, Washington, DC
  • Downie, J.A., Knight, C.D., Johnston, A.W.B., Rossen, L., Identification of genes and gene-products involved in the nodulation of peas by Rhizobium leguminosarum (1985) Mol. Gen. Genet, 198, pp. 255-262
  • Finnie, C., Hartley, N.M., Findlay, K.C., Downie, J.A., The Rhizobium leguminosarum prsDE genes are required for secretion of several proteins, some of which influence nodulation, symbiotic nitrogen fixation and exopolysaccharide modification (1997) Mol. Microbiol, 25, pp. 135-146
  • Finnie, C., Zorreguieta, A., Hartley, N.M., Downie, J.A., Characterization of Rhizobium leguminosarum exopolysaccharide glycanases that are secreted via a type I exporter and have a novel heptapeptide repeat motif (1998) J. Bacteriol, 180, pp. 1691-1699
  • Glenn, S.A., Gurich, N., Feeney, M.A., Gonzalez, J.E., The ExpR/Sin quorum-sensing system controls succinoglycan production in Sinorhizobium meliloti (2007) J. Bacteriol, 189, pp. 7077-7088
  • Gonzalez, N., Heeb, S., Valverde, C., Kay, E., Reimmann, C., Junier, T., Haas, D., Genome-wide search reveals a novel GacA-regulated small RNA in Pseudomonas species (2008) BMC Genomics, 9, p. 167
  • Gonzalez, V., Santamaria, R.I., Bustos, P., Hernandez-Gonzalez, I., Medrano-Soto, A., Moreno-Hagelsieb, G., Janga, S.C., Davila, G., The partitioned Rhizo- bium etli genome: Genetic and metabolic redundancy in seven interacting replicons (2006) Proc. Natl. Acad. Sci. USA, 103, pp. 3834-3839
  • Gray, K.M., Pearson, J.P., Downie, J.A., Boboye, B.E., Greenberg, E.P., Cell-to-cell signaling in the symbiotic nitrogen-fixing bacterium Rhizo- bium leguminosarum: Autoinduction of a stationary phase and rhizosphere- expressed genes (1996) J. Bacteriol, 178, pp. 372-376
  • Hammer, B.K., Bassler, B.L., Regulatorysmall RNAs circumvent the conventional quorum sensing pathway in pandemic Vibrio cholerae (2007) Proc. Natl. Acad. Sci. USA, 104, pp. 11145-11149
  • Heeb, S., Kuehne, S.A., Bycroft, M., Crivii, S., Allen, M.D., Haas, D., Camara, M., Williams, P., Functional analysis of the post-transcrip- tional regulator RsmA reveals a novel RNA-binding site (2006) J. Mol. Biol, 355, pp. 1026-1036
  • Hoang, H.H., Becker, A., Gonzalez, J.E., The LuxR homolog ExpR, in combination with the Sin quorum sensing system, plays a central role in Sinorhizobium meliloti gene expression (2004) J. Bacteriol, 186, pp. 5460-5472
  • Hoang, H.H., Gurich, N., Gonzaalez, J.E., Regulationofmotilityby the ExpR/Sin quorum-sensing system in Sinorhizobium meliloti (2008) J. Bacteriol, 190, pp. 861-871
  • Knight, C.D., Rossen, L., Robertson, J.G., Wells, B., Downie, J.A., Nodulation inhibition by Rhizobium leguminosarum multicopy nodABC genes and analysis of early stages of plant infection (1986) J. Bacteriol, 166, pp. 552-558
  • Kobayashi, K., Gradual activation of the response regulator DegU controls serial expression of genes for flagellum formation and biofilm formation in Bacillus subtilis (2007) Mol. Microbiol, 66, pp. 395-409
  • Kovach, M.E., Elzer, P.H., Hill, D.S., Robertson, G.T., Farris, M.A., Roop, R.M., Peterson, K.M., Four new derivatives of the broad-host-range cloning vector pBBR1mcs, carrying different antibiotic-resistance cassettes (1995) Gene, 166, pp. 175-176
  • Lamb, J.W., Downie, J.A., Johnston, A.W., Cloning of the nodulation (nod) genes of Rhizobium phaseoli and their homology to R. leguminosarum nod DNA (1985) Gene, 34, pp. 235-241
  • Lamb, J.W., Hombrecher, G., Johnston, A.W.B., Plasmid-determined nodulation and nitrogen-fixation abilities in Rhizobium phaseoli (1982) Mol. Gen. Genet, 186, pp. 449-452
  • Lenz, D.H., Bassler, B.L., The small nucleoid protein Fis is involved in Vibrio cholerae quorum sensing (2007) Mol. Microbiol, 63, pp. 859-871
  • Lithgow, J.K., Wilkinson, A., Hardman, A., Rodelas, B., Wisniewski-Dye, F., Williams, P., Downie, J.A., The regulatory locus cinRI in Rhizobium leguminosarum controls a network of quorum-sensing loci (2000) Mol. Microbiol, 37, pp. 81-97
  • Marketon, M.M., Glenn, S.A., Eberhard, A., Gonzalez, J.E., Quorum sensing controls exopolysaccharide production in Sinorhizobium meliloti (2003) J. Bacteriol, 185, pp. 325-331
  • McClean, K.H., Winson, M.K., Fish, L., Taylor, A., Chhabra, S.R., Camara, M., Daykin, M., Williams, P., Quorum sensing and Chromobacterium violaceum: Exploitation of violacein production and inhibition for the detection of N-acylho- moserine lactones (1997) Microbiology, 143, pp. 3703-3711
  • McIntosh, M., Krol, E., Becker, A., Competitive and cooperative effects in quorum-sensing-regulated galactoglucan biosynthesis in Sinorhizo- bium meliloti (2008) J. Bacteriol, 190, pp. 5308-5317
  • Pellock, B.J., Teplitski, M., Boinay, R.P., Bauer, W.D., Walker, G.C., A LuxR homolog controls production of symbiotically active extracellular polysaccharide II by Sinorhizobium meliloti (2002) J. Bacteriol, 184, pp. 5067-5076
  • Prentki, P., Krisch, H.M., In vitro insertional mutagenesis with a selectable DNA fragment (1984) Gene, 29, pp. 303-313
  • Quandt, J., Hynes, M.F., Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria (1993) Gene, 127, pp. 15-21
  • Rodelas, B., Lithgow, J.K., Wisniewski-Dye, F., Hardman, A., Wilkinson, A., Economou, A., Williams, P., Downie, J.A., Analysis ofquorum-sensing- dependent control of rhizosphere-expressed (rhi) genes in Rhizobium leggu- minosarum bv. viciae (1999) J. Bacteriol, 181, pp. 3816-3823
  • Rosemeyer, V., Michiels, J., Verreth, C., Vanderleyden, J., luxl- and luxR-homologous genes of Rhizobium etli CNPAF512 contribute to synthesis of autoinducer molecules and nodulation of Phaseolus vulgaris (1998) J. Bacteriol, 180, pp. 815-821
  • Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular cloning: A laboratory manual, , 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  • Sharma, S.B., Signer, E.R., Temporal and spatial regulation ofthe symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA (1990) Genes Dev, 4, pp. 344-356
  • Shaw, P.D., Ping, G., Daly, S.L., Cha, C., Cronan Jr., J.E., Rinehart, K.L., Farrand, S.K., Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography (1997) Proc. Natl. Acad. Sci. USA, 94, pp. 6036-6041
  • Sherwood, M.T., Improved synthetic medium for the growth of Rhi- zobium (1970) J. Appl. Bacteriol, 33, pp. 708-713
  • Sourjik, V., Muschler, P., Scharf, B., Schmitt, R., VisN and VisR are global regulators of chemotaxis, flagellar, and motility genes in Sinorhizo- bium (Rhizobium) meliloti (2000) J. Bacteriol, 182, pp. 782-788
  • Spaink, H.P., Wijffelman, C.A., Pees, E., Okker, R.J.H., Lugtenberg, B.J.J., Rhizobium nodulation gene nodD as a determinant of host specificity (1987) Nature, 328, pp. 337-340
  • Svenningsen, S.L., Waters, C.M., Bassler, B.L., A negative feedback loop involving small RNAs accelerates Vibrio cholerae transition out of quorum-sensing mode (2008) Genes Dev, 22, pp. 226-238
  • Thorne, S.H., Williams, H.D., Cell density-dependent starvation survival of Rhizobium leguminosarum bv. phaseoli: Identification of the role of an N-acyl homoserine lactone in adaptation to stationary-phase survival (1999) J. Bacteriol, 181, pp. 981-990
  • Tu, K.C., Bassler, B.L., Multiple small RNAs act additively to integrate sensory information and control quorum sensing in Vibrio harveyi (2007) Genes Dev, 21, pp. 221-233
  • Whitehead, N.A., Barnard, A.M., Slater, H., Simpson, N.J., Salmond, G.P., Quorum-sensing in Gram-negative bacteria (2001) FEMS Micro- biol. Rev, 25, pp. 365-404
  • Wilkinson, A., Danino, V., Wisniewski-Dye, F., Lithgow, J.K., Downie, J.A., N-acyl-homoserine lactone inhibition of rhizobial growth is mediated by two quorum-sensing genes that regulate plasmid transfer (2002) J. Bacteriol, 184, pp. 4510-4519
  • Winans, S.C., Cell-cell signalling within crown gall tumors (2008) Chemical communication among bacteria, pp. 291-306. , S. C. Winans and B. L. Bassler ed, ASM Press, Washington, DC
  • Wisniewski-Dye, F., Downie, J.A., Quorum-sensing in Rhizobium (2002) Antonie van Leeuwenhoek, 81, pp. 397-407
  • Wisniewski-Dyae, F., Jones, J., Chhabra, S.R., Downie, J.A., raiIR genes are part of a quorum-sensing network controlled by cinI and cinR in Rhizobium leguminosarum (2002) J. Bacteriol, 184, pp. 1597-1606
  • Young, J. P. W., L. C. Crossman, A. W. B. Johnston, N. R. Thomson, Z. F. Ghazoui, K. H. Hull, M. Wexler, A. R. J. Curson, J. D. Todd, P. S. Poole, T. H. Mauchline, A. K. East, M. A. Quail, C. Churcher, C. Arrowsmith, I. Cherevach, T. Chillingworth, K. Clarke, A. Cronin, P. Davis, A. Fraser, Z. Hance, H. Hauser, K. Jagels, S. Moule, K. Mungall, H. Norbertczak, E. Rabbinowitsch, M. Sanders, M. Simmonds, S. Whitehead, and J. Parkhill. 2006. The genome of Rhizobium leguminosarum has recognizable core and accessory components. Genome Biology 7:R34; Zhu, J., Oger, P.M., Schrammeijer, B., Hooykaas, P.J., Farrand, S.K., Winans, S.C., The bases of crown gall tumorigenesis (2000) J. Bacteriol, 182, pp. 3885-3895
  • Zorreguieta, A., Finnie, C., Downie, J.A., Extracellular glycanases of Rhizobium leguminosarum are activated on the cell surface by an exopo- lysaccharide-related component (2000) J. Bacteriol, 182, pp. 1304-1312


---------- APA ----------
Edwards, A., Frederix, M., Wisniewski-Dyé, F., Jones, J., Zorreguieta, A. & Allan Downie, J. (2009) . The cin and rai quorum-sensing regulatory systems in Rhizobium leguminosarum are coordinated by ExpR and CinS, a small regulatory protein coexpressed with CinI. Journal of Bacteriology, 191(9), 3059-3067.
---------- CHICAGO ----------
Edwards, A., Frederix, M., Wisniewski-Dyé, F., Jones, J., Zorreguieta, A., Allan Downie, J. "The cin and rai quorum-sensing regulatory systems in Rhizobium leguminosarum are coordinated by ExpR and CinS, a small regulatory protein coexpressed with CinI" . Journal of Bacteriology 191, no. 9 (2009) : 3059-3067.
---------- MLA ----------
Edwards, A., Frederix, M., Wisniewski-Dyé, F., Jones, J., Zorreguieta, A., Allan Downie, J. "The cin and rai quorum-sensing regulatory systems in Rhizobium leguminosarum are coordinated by ExpR and CinS, a small regulatory protein coexpressed with CinI" . Journal of Bacteriology, vol. 191, no. 9, 2009, pp. 3059-3067.
---------- VANCOUVER ----------
Edwards, A., Frederix, M., Wisniewski-Dyé, F., Jones, J., Zorreguieta, A., Allan Downie, J. The cin and rai quorum-sensing regulatory systems in Rhizobium leguminosarum are coordinated by ExpR and CinS, a small regulatory protein coexpressed with CinI. J. Bacteriol. 2009;191(9):3059-3067.