1887

Abstract

is a nitrogen-fixing soil bacterium that undergoes differentiation to form cysts resistant to desiccation. Upon encystment, this bacterium becomes non-motile. As in enteric bacteria, motility in occurs through the use of peritrichous flagella. , a phylogenetically close relative of , possesses a single polar flagellum. The FlhDC proteins are the master regulators of flagella and motility in enterobacteria, whereas FleQ is the master regulator in , and it is under AlgU (sigmaE) negative control. At present, nothing is known about the organization and expression of flagella genes in . Here, we identified the flagella gene cluster of this bacterium. Homologues of the master regulatory genes and are present in . Inactivation of , but not , impaired flagella biogenesis and motility. We present evidence indicating that a negative effect of the AlgU sigma factor on expression causes loss of motility in , and that CydR (a homologue of Fnr) is under AlgU control and has a negative effect on expression. Taken together, these results suggest the existence of a cascade consisting of AlgU and CydR that negatively controls expression of ; the results also suggest that the block in flagella synthesis under encystment conditions centres on repression by the AlgU–CydR cascade.

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2008-06-01
2024-03-28
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References

  1. Aldridge P., Hughes K. T. 2002; Regulation of flagellar assembly. Curr Opin Microbiol 5:160–165
    [Google Scholar]
  2. Alexeyev M. F., Shokolenko I., Croughan T. P. 1995; Improved antibiotic-resistance gene cassettes and omega elements for Escherichia coli vector construction and in vitro deletion/insertion mutagenesis. Gene 160:63–67
    [Google Scholar]
  3. Arora S. K., Ritchings B. W., Almira E. C., Lory S., Ramphal R. 1997; A transcriptional activator, FleQ, regulates mucin adhesion and flagellar gene expression in Pseudomonas aeruginosa in a cascade manner. J Bacteriol 179:5574–5581
    [Google Scholar]
  4. Bali A., Blanco G., Hill S., Kennedy C. 1992; Excretion of ammonium by a nifL mutant of Azotobacter vinelandii fixing nitrogen. Appl Environ Microbiol 58:1711–1718
    [Google Scholar]
  5. Blatny J. M., Brautaset T., Winther-Larsen H. C., Haugan K., Valla S. 1997; Construction and use of a versatile set of broad-host-range cloning and expression vectors based on the RK2 replicon. Appl Environ Microbiol 63:370–374
    [Google Scholar]
  6. Campos M.-E., Martínez-Salazar J. M., Lloret L., Moreno S., Núñez C., Espín G., Soberón-Chávez G. 1996; Characterization of the gene coding for GDP-mannose dehydrogenase ( algD ) from Azotobacter vinelandii . J Bacteriol 178:1793–1799
    [Google Scholar]
  7. Clarke M. B., Sperandio V. 2005; Transcriptional regulation of flhDC by QseBC and sigma (FliA) in enterohaemorragic Escherichia coli . Mol Microbiol 57:1734–1749
    [Google Scholar]
  8. Dasgupta N., Ramphal R. 2001; Interaction of the antiactivator FleN with the transcriptional activator FleQ regulates flagellar number in Pseudomonas aeruginosa . J Bacteriol 183:6636–6644
    [Google Scholar]
  9. Dasgupta N., Wolfgang M. C., Goodman A. L., Arora S. K., Jyot J., Lory S., Ramphal R. 2003; A four-tiered transcriptional regulatory circuit controls flagellar biogenesis in Pseudomonas aeruginosa . Mol Microbiol 50:809–824
    [Google Scholar]
  10. Fellay R., Frey J., Krisch H. 1987; Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis. Gene 52:147–154
    [Google Scholar]
  11. Francez-Charlot A., Laugel B., Van Gemert A., Dubarry N., Wiorowski F., Castanie-Cornet M. P., Gutierrez C., Cam K. 2003; RcsCDB His-Asp phosphorelay system negatively regulates the flhDC operon in Escherichia coli . Mol Microbiol 49:823–832
    [Google Scholar]
  12. Gaona G., Núñez C., Goldberg J. B., Linford A. N., Nájera R., Castañeda M., Guzmán J., Espín G., Soberón-Chávez G. 2004; Characterization of the Azotobacter vinelandii algC gene involved in alginate and lipopolysaccharide production. FEMS Microbiol Lett 238:199–206
    [Google Scholar]
  13. Garrett E. S., Perlegas D., Wozniak D. J. 1999; Negative control of flagellum synthesis in Pseudomonas aeruginosa is modulated by the alternative sigma factor AlgT (AlgU. J Bacteriol 181:7401–7404
    [Google Scholar]
  14. Hedges R. W., Baumberg S. 1973; Resistance to arsenic compounds conferred by a plasmid transmissible between strains of Escherichia coli . J Bacteriol 115:459–460
    [Google Scholar]
  15. Hitchins V. M., Sadoff H. L. 1973; Sequential metabolic events during encystment in Azotobacter vinelandii . J Bacteriol 113:1273–1279
    [Google Scholar]
  16. Holt J. G., Krieg N. R., Sneath P. H. A., Staley J. T., Williams S. T. 1994 Bergey's Manual of Determinative Bacteriology , 9th edn. p 79 Baltimore: Williams & Wilkins;
    [Google Scholar]
  17. Jyot J., Dasgupta N., Ramphal R. 2002; FleQ, the major flagellar gene regulator in Pseudomonas aeruginosa , binds to enhancer sites located either upstream or atypically downstream of the RpoN binding site. J Bacteriol 184:5251–5260
    [Google Scholar]
  18. Kennedy C., Gamal R., Hummprey R., Ramos J., Brigle K., Dean D. 1986; The nifH , nifM , and nifN genes of Azotobacter vinelandii : characterization by Tn 5 mutagenesis and isolation from pLARF1 gene banks. Mol Gen Genet 205:318–325
    [Google Scholar]
  19. Kutsukake K. 1997; Autogenous and global control of the flagellar master operon, flhDC , in Salmonella typhimurium . Mol Gen Genet 254:440–448
    [Google Scholar]
  20. Kutsukake K., Nambu T. 2000; Bacterial flagellum: a paradigm for biogenesis of transenvelope supramolecular structures. Recent Res Dev Microbiol 4:607–615
    [Google Scholar]
  21. Macnab R. M. others 1996; Flagella and motility. In Escherichia coli and Salmonella: Cellular and Molecular Biology pp 123–145 Edited by Neidhardt F. C. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  22. Martin D. W., Schurr M. J., Mudd M. H., Govan J. R., Holloway B. W., Deretic V. 1993; Mechanism of conversion to mucoidy in P. aeruginosa infecting cystic fibrosis patients. Proc Natl Acad Sci U S A 90:8377–8381
    [Google Scholar]
  23. Martínez-Salazar J. M., Moreno S., Nájera R., Boucher J. C., Espín G., Soberón-Chávez G., Deretic V. 1996; Characterization of the genes coding for the putative sigma factor AlgU and its regulators MucA, MucB, MucC, and MucD in Azotobacter vinelandii and evaluation of their roles in alginate biosynthesis. J Bacteriol 178:1800–1808
    [Google Scholar]
  24. Mathee K., McPherson C. J., Ohman D. E. 1997; Posttranslational control of the AlgT ( algU )-encoded sigma22 for expression of the alginate regulon in Pseudomonas aeruginosa and localization of its antagonist proteins MucA and MucB (AlgN. J Bacteriol 179:3711–3720
    [Google Scholar]
  25. Mejía-Ruíz H., Moreno S., Guzmán J., Nájera R., León R., Soberón-Chávez G., Espín G. 1997; Isolation and characterization of an Azotobacter vinelandii algK mutant. FEMS Microbiol Lett 156:101–106
    [Google Scholar]
  26. Miller J. H. 1972 Experiments in Molecular Genetics pp 431–435 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  27. Moreno S., Nájera R., Guzmán J., Soberón-Chávez G., Espín G. 1998; Role of alternative factor AlgU in encystment of Azotobacter vinelandii . J Bacteriol 180:2766–2769
    [Google Scholar]
  28. Noguez R., Segura D., Moreno S., Hernandez A., Juárez K., Espín G. 2008; Enzyme INtr, Npr, and IIANtr are involved in regulation of the polyhydroxybutyrate biosynthetic genes in Azotobacter vinelandii . J Mol Microbiol Biotechnol in press
    [Google Scholar]
  29. Núñez C., León R., Guzmán J., Espín G., Soberón-Chávez G. 2000; Role of Azotobacter vinelandii mucA and mucC gene products in alginate production. J Bacteriol 182:6550–6556
    [Google Scholar]
  30. Page W. J., Sadoff H. L. 1975; Relationship between calcium and uronic acids in the encystment of Azotobacter vinelandii . J Bacteriol 122:145–151
    [Google Scholar]
  31. Page W. J., von Tigerstrom M. 1978; Induction of transformation competence in Azotobacter vinelandii iron limited cultures. Can J Microbiol 24:1590–1594
    [Google Scholar]
  32. Peralta-Gíl M., Segura D., Guzmán J., Servín-Gonzalez L., Espín G. 2002; Expression of the Azotobacter vinelandii poly- β -hydroxybutyrate biosynthetic phbBAC operon is driven by two overlapping promoters and is dependent on the transcriptional activator PhbR. J Bacteriol 184:5672–5677
    [Google Scholar]
  33. Qiu D., Eisinger V. M., Rowen D. W., Yu H. D. 2007; Regulated proteolysis controls mucoid conversion in Pseudomonas aeruginosa . Proc Natl Acad Sci U S A 104:8107–8112
    [Google Scholar]
  34. Ramsey D. M., Wosniak D. J. 2005; Understanding the control of Pseudomonas aeruginosa alginate synthesis and the prospects for management of chronic infections in cystic fibrosis. Mol Microbiol 56:309–322
    [Google Scholar]
  35. Rediers H., Vanderleyden J., De Mot R. 2004; Azotobacter vinelandii : a Pseudomonas in disguise?. Microbiology 150:1117–1119
    [Google Scholar]
  36. Ritchings B. W., Almira E. C., Lory S., Ramphal R. 1995; Cloning and phenotypic characterization of fleS and fleR , new response regulators of Pseudomonas aeruginosa which regulate motility and adhesion to mucin. Infect Immun 63:4868–4876
    [Google Scholar]
  37. Sadoff H. L. 1975; Encystment and germination in Azotobacter vinelandii . Bacteriol Rev 39:516–539
    [Google Scholar]
  38. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  39. Schurr M. J., Yu H., Martinez-Salazar J. M., Boucher J. C., Deretic V. 1996; Control of AlgU, a member of the sigma E-like family of stress sigma factors, by the negative regulators MucA and MucB and Pseudomonas aeruginosa conversion to mucoidy in cystic fibrosis. J Bacteriol 178:4997–5004
    [Google Scholar]
  40. Soutourina O., Kolb A., Krin E., Laurent-Winter C., Rimsky S., Danchin A., Bertin P. 1999; Multiple control of flagellum biosynthesis in Escherichia coli : role of H-NS protein and the cyclic AMP-catabolite activator protein complex in transcription of the flhDC master operon. J Bacteriol 181:7500–7508
    [Google Scholar]
  41. Tart A. H., Wolfgang M. C., Wosniak D. J. 2005; The alternative sigma factor AlgT represses Pseudomonas aeruginosa flagellum biosynthesis by inhibiting expression of fleQ . J Bacteriol 187:7955–7962
    [Google Scholar]
  42. Tart A. H., Blanks M. J., Wosniak D. J. 2006; The AlgT-dependent transcriptional regulator AmrZ (AlgZ) inhibits flagellum biosynthesis in mucoid, nonmotile Pseudomonas aeruginosa cystic fibrosis isolates. J Bacteriol 188:6483–6489
    [Google Scholar]
  43. Wei B. L., Brun-Zinkernagel A. M., Simecka J. W., Pruss B. M., Babitzke P., Romeo T. 2001; Positive regulation of motility and flhDC expression by the RNA-binding protein CsrA of Escherichia coli . Mol Microbiol 40:245–256
    [Google Scholar]
  44. Wozniak D. J., Ohman D. E. 1994; Transcriptional analysis of the Pseudomonas aeruginosa genes algR , algB , and algD reveals a hierarchy of alginate gene expression which is modulated by algT . J Bacteriol 176:6007–6014
    [Google Scholar]
  45. Wu G., Cruz-Ramos H., Hill S., Green J., Sawers G., Poole R. K. 2000; Regulation of cytochrome bd expression in the obligate aerobe Azotobacter vinelandii by CydR (Fnr). Sensitivity to oxygen species, and nitric oxide. J Biol Chem 275:4679–4686
    [Google Scholar]
  46. Xie Z. D., Hershberger C. D., Shankar S., Ye R. W., Chakrabarty A. M. 1996; Sigma factor-anti-sigma factor interaction in alginate synthesis: inhibition of AlgT by MucA. J Bacteriol 178:4990–4996
    [Google Scholar]
  47. Yanagihara S., Iyoda S., Ohnishi K., Iino T., Kutsukake K. 1999; Structure and transcriptional control of the flagellar master operon of Salmonella typhimurium . Genes Genet Syst 74:105–111
    [Google Scholar]
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