Role of the GGDEF regulator PleD in polar development of Caulobacter crescentus

Several members of the two‐component signal transduction family have been implicated in the control of  polar development in Caulobacter crescentus: PleC and DivJ, two polarly localized histidine sensor kinases; and the response regulators DivK and PleD. The PleD protein was shown previously to be required during the swarmer‐to‐stalked cell transition for flagellar ejection and efficient stalk biogenesis. Here, we present data indicating that PleD also controls the onset of motility and a cell density switch immediately preceding cell division. Constitutively active alleles of pleD or wspR, an orthologue from Pseudomonas fluorescens, almost completely suppressed C. crescentus motility and inhibited the increase in swarmer cell density during cell differentiation. The observation that these alleles also had a dominant‐negative effect on motility in a pleC divJ and a pleC divK mutant background indicated that PleD is located downstream of the other components in the signal transduction cascade, which controls the activity of the flagellar motor. In addition, the presence of a constitutive pleD or wspR allele resulted in a doubling of the average stalk length. Together, this is consistent with a model in which the active form of PleD, PleD∼P, negatively controls aspects of differentiation in the late predivisional cell, whereas it acts positively on polar development during the swarmer‐to‐stalked cell transition. In agreement with such a model, we found that DivJ, which localizes to the stalked pole during cell differentiation, positively controlled the in vivo phosphorylation status of PleD, and the swarmer pole‐specific PleC kinase modulated this status in a negative manner. Furthermore, domain switch experiments demonstrated that the WspR GGDEF output domain from P. fluorescens is active in C. crescentus, favouring a more general function for this novel signalling domain over a specific role such as DNA or protein interaction. Possible roles for PleD and its C‐terminal output domain in modulating the polar cell surface of C. crescentus are discussed.

[1]  U. Jenal,et al.  The Caulobacter cell cycle: timing, spatial organization and checkpoints. , 2002, Current opinion in microbiology.

[2]  A. Pühler,et al.  A Broad Host Range Mobilization System for In Vivo Genetic Engineering: Transposon Mutagenesis in Gram Negative Bacteria , 1983, Bio/Technology.

[3]  N. Ausmees,et al.  Structural and putative regulatory genes involved in cellulose synthesis in Rhizobium leguminosarum bv. trifolii. , 1999, Microbiology.

[4]  Y. Bum,et al.  The Dimorphic Life Cycle of Caulobacter and Stalked Bacteria , 2000 .

[5]  E. G. Ninfa,et al.  A histidine protein kinase homologue required for regulation of bacterial cell division and differentiation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[6]  L. Shapiro,et al.  Differential localization of two histidine kinases controlling bacterial cell differentiation. , 1999, Molecular cell.

[7]  Michael Y. Galperin,et al.  Novel domains of the prokaryotic two-component signal transduction systems. , 2001, FEMS microbiology letters.

[8]  B Ely,et al.  A histidine protein kinase is involved in polar organelle development in Caulobacter crescentus. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[9]  N. Ohta,et al.  Signal transduction in the cell cycle regulation of Caulobacter differentiation. , 1996, Trends in microbiology.

[10]  M. Gilles-Gonzalez,et al.  Dos, a heme-binding PAS protein from Escherichia coli, is a direct oxygen sensor. , 2000, Biochemistry.

[11]  Lucy Shapiro,et al.  Cell Type-Specific Phosphorylation and Proteolysis of a Transcriptional Regulator Controls the G1-to-S Transition in a Bacterial Cell Cycle , 1997, Cell.

[12]  D. Amikam,et al.  The cyclic diguanylic acid regulatory system of cellulose synthesis in Acetobacter xylinum. Chemical synthesis and biological activity of cyclic nucleotide dimer, trimer, and phosphothioate derivatives. , 1990, The Journal of biological chemistry.

[13]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[14]  J Wu,et al.  A novel bacterial tyrosine kinase essential for cell division and differentiation. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Peter Ross,et al.  Three cdg Operons Control Cellular Turnover of Cyclic Di-GMP in Acetobacter xylinum: Genetic Organization and Occurrence of Conserved Domains in Isoenzymes , 1998, Journal of bacteriology.

[16]  A. Newton,et al.  Pseudoreversion analysis indicates a direct role of cell division genes in polar morphogenesis and differentiation in Caulobacter crescentus. , 1991, Genetics.

[17]  M. Travisano,et al.  Adaptive divergence in experimental populations of Pseudomonas fluorescens. I. Genetic and phenotypic bases of wrinkly spreader fitness. , 2002, Genetics.

[18]  B. Ely.  Genetics of Caulobacter crescentus. , 1991, Methods in enzymology.

[19]  M. Rohde,et al.  The multicellular morphotypes of Salmonella typhimurium and Escherichia coli produce cellulose as the second component of the extracellular matrix , 2001, Molecular microbiology.

[20]  A. Newton,et al.  An essential single domain response regulator required for normal cell division and differentiation in Caulobacter crescentus. , 1995, The EMBO journal.

[21]  C. Ponting,et al.  The GAF domain: an evolutionary link between diverse phototransducing proteins. , 1997, Trends in biochemical sciences.

[22]  R. Turner,et al.  Characterization of high density monolayers of the biofilm bacterium Caulobacter crescentus: evaluating prospects for developing immobilized cell bioreactors. , 2000, Canadian journal of microbiology.

[23]  D. Amikam,et al.  Genetic data indicate that proteins containing the GGDEF domain possess diguanylate cyclase activity. , 2001, FEMS microbiology letters.

[24]  M. Igo,et al.  Differential Expression of the OmpF and OmpC Porin Proteins in Escherichia coli K-12 Depends upon the Level of Active OmpR , 1998, Journal of bacteriology.

[25]  J. H. Boom,et al.  Regulation of cellulose synthesis in Acetobacter xylinum by cyclic diguanylic acid , 1987, Nature.

[26]  L. Shapiro,et al.  Dynamic localization of a cytoplasmic signal transduction response regulator controls morphogenesis during the Caulobacter cell cycle , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Osteras,et al.  Regulatory circuits in Caulobacter. , 2000, Current opinion in microbiology.

[28]  M. Saier,et al.  Response regulators of bacterial signal transduction systems: Selective domain shuffling during evolution , 1995, Journal of Molecular Evolution.

[29]  A. Newton,et al.  An essential, multicomponent signal transduction pathway required for cell cycle regulation in Caulobacter. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Kolter,et al.  Biofilm formation as microbial development. , 2000, Annual review of microbiology.

[31]  H. Sambrook.  Molecular cloning : a laboratory manual. Cold Spring Harbor, NY , 1989 .

[32]  Michael Y. Galperin,et al.  MHYT, a new integral membrane sensor domain. , 2001, FEMS microbiology letters.

[33]  N. Agabian,et al.  Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells , 1977, Journal of bacteriology.

[34]  I. Zhulin,et al.  PAS domain S-boxes in Archaea, Bacteria and sensors for oxygen and redox. , 1997, Trends in biochemical sciences.

[35]  Roles of the histidine protein kinase pleC in Caulobacter crescentus motility and chemotaxis , 1997, Journal of bacteriology.

[36]  D. Durocher,et al.  The FHA domain is a modular phosphopeptide recognition motif. , 1999, Molecular cell.

[37]  A. Newton,et al.  Identification of a novel response regulator required for the swarmer-to-stalked-cell transition in Caulobacter crescentus , 1995, Journal of bacteriology.

[38]  R Mayer,et al.  Cellulose biosynthesis and function in bacteria. , 1991, Microbiological reviews.

[39]  N. Grishin,et al.  GGDEF domain is homologous to adenylyl cyclase , 2001, Proteins.

[40]  David A. D'Argenio,et al.  Autolysis and Autoaggregation in Pseudomonas aeruginosa Colony Morphology Mutants , 2002, Journal of bacteriology.

[41]  H. McAdams,et al.  Global analysis of the genetic network controlling a bacterial cell cycle. , 2000, Science.

[42]  C. Yanisch-Perron,et al.  Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. , 1985, Gene.

[43]  R. Schmitt,et al.  Different roles of CheY1 and CheY2 in the chemotaxis of Rhizobium meliloti , 1996, Molecular microbiology.

[44]  D. Kaiser,et al.  The act operon controls the level and time of C‐signal production for Myxococcus xanthus development , 2001, Molecular microbiology.

[45]  M H Saier,et al.  Structural, functional, and evolutionary relationships among extracellular solute-binding receptors of bacteria , 1993, Microbiological reviews.

[46]  C. Dieffenbach,et al.  PCR primer: a laboratory manual. , 2003 .

[47]  S. Kustu,et al.  Glutamate at the site of phosphorylation of nitrogen-regulatory protein NTRC mimics aspartyl-phosphate and activates the protein. , 1993, Journal of molecular biology.

[48]  A. Newton,et al.  Turning off flagellum rotation requires the pleiotropic gene pleD: pleA, pleC, and pleD define two morphogenic pathways in Caulobacter crescentus , 1989, Journal of bacteriology.

[49]  S. Normark,et al.  AgfD, the checkpoint of multicellular and aggregative behaviour in Salmonella typhimurium regulates at least two independent pathways , 2000, Molecular microbiology.

[50]  C. Ponting,et al.  PAS: a multifunctional domain family comes to light , 1997, Current Biology.

[51]  U. Jenal,et al.  Cell cycle‐dependent degradation of a flagellar motor component requires a novel‐type response regulator , 1999, Molecular microbiology.