Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Sep;179(17):5502–5510. doi: 10.1128/jb.179.17.5502-5510.1997

A gene coding for a putative sigma 54 activator is developmentally regulated in Caulobacter crescentus.

M V Marques 1, S L Gomes 1, J W Gober 1
PMCID: PMC179422  PMID: 9287006

Abstract

In Caulobacter crescentus, the alternative sigma factor sigma54 plays an important role in the expression of late flagellar genes. Sigma54-dependent genes are temporally and spatially controlled, being expressed only in the swarmer pole of the predivisional cell. The only sigma54 activator described so far is the FlbD protein, which is involved in activation of the class III and IV flagellar genes and repression of the fliF promoter. To identify new roles for sigma54 in the metabolism and differentiation of C. crescentus, we cloned and characterized a gene encoding a putative sigma54 activator, named tacA. The deduced amino acid sequence from tacA has high similarity to the proteins from the NtrC family of transcriptional activators, including the aspartate residues that are phosphorylated by histidine kinases in other activators. The promoter region of the tacA gene contains a conserved sequence element present in the promoters of class II flagellar genes, and tacA shows a temporal pattern of expression similar to the patterns of these genes. We constructed an insertional mutant that is disrupted in tacA (strain SP2016), and an analysis of this strain showed that it has all polar structures, such as pili, stalk, and flagellum, and displays a motile phenotype, indicating that tacA is not involved in the flagellar biogenesis pathway. However, this strain has a high percentage of filamentous cells and shows a clear-plaque phenotype when infected with phage phiCb5. These results suggest that the TacA protein could mediate the effect of sigma54 on a different pathway in C. crescentus.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agabian-Keshishian N., Shapiro L. Stalked bacteria: properties of deoxriybonucleic acid bacteriophage phiCbK. J Virol. 1970 Jun;5(6):795–800. doi: 10.1128/jvi.5.6.795-800.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alley M. R., Gomes S. L., Alexander W., Shapiro L. Genetic analysis of a temporally transcribed chemotaxis gene cluster in Caulobacter crescentus. Genetics. 1991 Oct;129(2):333–341. doi: 10.1093/genetics/129.2.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson D. K., Ohta N., Wu J., Newton A. Regulation of the Caulobacter crescentus rpoN gene and function of the purified sigma 54 in flagellar gene transcription. Mol Gen Genet. 1995 Mar 20;246(6):697–706. doi: 10.1007/BF00290715. [DOI] [PubMed] [Google Scholar]
  4. Bender R. A., Refson C. M., O'Neill E. A. Role of the flagellum in cell-cycle-dependent expression of bacteriophage receptor activity in Caulobacter crescentus. J Bacteriol. 1989 Feb;171(2):1035–1040. doi: 10.1128/jb.171.2.1035-1040.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bendis I., Shapiro L. Properties of Caulobacter ribonucleic acid bacteriophage phi Cb5. J Virol. 1970 Dec;6(6):847–854. doi: 10.1128/jvi.6.6.847-854.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benson A. K., Ramakrishnan G., Ohta N., Feng J., Ninfa A. J., Newton A. The Caulobacter crescentus FlbD protein acts at ftr sequence elements both to activate and to repress transcription of cell cycle-regulated flagellar genes. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4989–4993. doi: 10.1073/pnas.91.11.4989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brun Y. V., Marczynski G., Shapiro L. The expression of asymmetry during Caulobacter cell differentiation. Annu Rev Biochem. 1994;63:419–450. doi: 10.1146/annurev.bi.63.070194.002223. [DOI] [PubMed] [Google Scholar]
  8. Brun Y. V., Shapiro L. A temporally controlled sigma-factor is required for polar morphogenesis and normal cell division in Caulobacter. Genes Dev. 1992 Dec;6(12A):2395–2408. doi: 10.1101/gad.6.12a.2395. [DOI] [PubMed] [Google Scholar]
  9. Buikema W. J., Szeto W. W., Lemley P. V., Orme-Johnson W. H., Ausubel F. M. Nitrogen fixation specific regulatory genes of Klebsiella pneumoniae and Rhizobium meliloti share homology with the general nitrogen regulatory gene ntrC of K. pneumoniae. Nucleic Acids Res. 1985 Jun 25;13(12):4539–4555. doi: 10.1093/nar/13.12.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chen W. P., Kuo T. T. A simple and rapid method for the preparation of gram-negative bacterial genomic DNA. Nucleic Acids Res. 1993 May 11;21(9):2260–2260. doi: 10.1093/nar/21.9.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chopra A. K., Peterson J. W., Prasad R. Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium. Biochim Biophys Acta. 1991 Dec 2;1129(1):115–118. doi: 10.1016/0167-4781(91)90224-a. [DOI] [PubMed] [Google Scholar]
  12. Dingwall A., Garman J. D., Shapiro L. Organization and ordered expression of Caulobacter genes encoding flagellar basal body rod and ring proteins. J Mol Biol. 1992 Dec 20;228(4):1147–1162. doi: 10.1016/0022-2836(92)90322-b. [DOI] [PubMed] [Google Scholar]
  13. Dingwall A., Zhuang W. Y., Quon K., Shapiro L. Expression of an early gene in the flagellar regulatory hierarchy is sensitive to an interruption in DNA replication. J Bacteriol. 1992 Mar;174(6):1760–1768. doi: 10.1128/jb.174.6.1760-1768.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ely B., Ely T. W. Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus. Genetics. 1989 Dec;123(4):649–654. doi: 10.1093/genetics/123.4.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ely B. Genetics of Caulobacter crescentus. Methods Enzymol. 1991;204:372–384. doi: 10.1016/0076-6879(91)04019-k. [DOI] [PubMed] [Google Scholar]
  16. Errington J. Bacillus subtilis sporulation: regulation of gene expression and control of morphogenesis. Microbiol Rev. 1993 Mar;57(1):1–33. doi: 10.1128/mr.57.1.1-33.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Evinger M., Agabian N. Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells. J Bacteriol. 1977 Oct;132(1):294–301. doi: 10.1128/jb.132.1.294-301.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  19. Foster-Hartnett D., Cullen P. J., Monika E. M., Kranz R. G. A new type of NtrC transcriptional activator. J Bacteriol. 1994 Oct;176(20):6175–6187. doi: 10.1128/jb.176.20.6175-6187.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gober J. W., Boyd C. H., Jarvis M., Mangan E. K., Rizzo M. F., Wingrove J. A. Temporal and spatial regulation of fliP, an early flagellar gene of Caulobacter crescentus that is required for motility and normal cell division. J Bacteriol. 1995 Jul;177(13):3656–3667. doi: 10.1128/jb.177.13.3656-3667.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gober J. W., Champer R., Reuter S., Shapiro L. Expression of positional information during cell differentiation of Caulobacter. Cell. 1991 Jan 25;64(2):381–391. doi: 10.1016/0092-8674(91)90646-g. [DOI] [PubMed] [Google Scholar]
  22. Gober J. W., Marques M. V. Regulation of cellular differentiation in Caulobacter crescentus. Microbiol Rev. 1995 Mar;59(1):31–47. doi: 10.1128/mr.59.1.31-47.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gober J. W., Shapiro L. A developmentally regulated Caulobacter flagellar promoter is activated by 3' enhancer and IHF binding elements. Mol Biol Cell. 1992 Aug;3(8):913–926. doi: 10.1091/mbc.3.8.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Gomes S. L., Shapiro L. Differential expression and positioning of chemotaxis methylation proteins in Caulobacter. J Mol Biol. 1984 Sep 25;178(3):551–568. doi: 10.1016/0022-2836(84)90238-9. [DOI] [PubMed] [Google Scholar]
  25. Grimm C., Panopoulos N. J. The predicted protein product of a pathogenicity locus from Pseudomonas syringae pv. phaseolicola is homologous to a highly conserved domain of several procaryotic regulatory proteins. J Bacteriol. 1989 Sep;171(9):5031–5038. doi: 10.1128/jb.171.9.5031-5038.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  27. Inouye S., Nakazawa A., Nakazawa T. Nucleotide sequence of the regulatory gene xylR of the TOL plasmid from Pseudomonas putida. Gene. 1988 Jun 30;66(2):301–306. doi: 10.1016/0378-1119(88)90366-6. [DOI] [PubMed] [Google Scholar]
  28. Ishimoto K. S., Lory S. Identification of pilR, which encodes a transcriptional activator of the Pseudomonas aeruginosa pilin gene. J Bacteriol. 1992 Jun;174(11):3514–3521. doi: 10.1128/jb.174.11.3514-3521.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jenal U., White J., Shapiro L. Caulobacter flagellar function, but not assembly, requires FliL, a non-polarly localized membrane protein present in all cell types. J Mol Biol. 1994 Oct 21;243(2):227–244. doi: 10.1006/jmbi.1994.1650. [DOI] [PubMed] [Google Scholar]
  30. Lagenaur C., Agabian N. Caulobacter crescentus pili: structure and stage-specific expression. J Bacteriol. 1977 Jul;131(1):340–346. doi: 10.1128/jb.131.1.340-346.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McCleary W. R., Zusman D. R. Purification and characterization of the Myxococcus xanthus FrzE protein shows that it has autophosphorylation activity. J Bacteriol. 1990 Dec;172(12):6661–6668. doi: 10.1128/jb.172.12.6661-6668.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Morett E., Segovia L. The sigma 54 bacterial enhancer-binding protein family: mechanism of action and phylogenetic relationship of their functional domains. J Bacteriol. 1993 Oct;175(19):6067–6074. doi: 10.1128/jb.175.19.6067-6074.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mullin D. A., Van Way S. M., Blankenship C. A., Mullin A. H. FlbD has a DNA-binding activity near its carboxy terminus that recognizes ftr sequences involved in positive and negative regulation of flagellar gene transcription in Caulobacter crescentus. J Bacteriol. 1994 Oct;176(19):5971–5981. doi: 10.1128/jb.176.19.5971-5981.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Newton A., Ohta N., Ramakrishnan G., Mullin D., Raymond G. Genetic switching in the flagellar gene hierarchy of Caulobacter requires negative as well as positive regulation of transcription. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6651–6655. doi: 10.1073/pnas.86.17.6651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. North A. K., Klose K. E., Stedman K. M., Kustu S. Prokaryotic enhancer-binding proteins reflect eukaryote-like modularity: the puzzle of nitrogen regulatory protein C. J Bacteriol. 1993 Jul;175(14):4267–4273. doi: 10.1128/jb.175.14.4267-4273.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Popham D. L., Szeto D., Keener J., Kustu S. Function of a bacterial activator protein that binds to transcriptional enhancers. Science. 1989 Feb 3;243(4891):629–635. doi: 10.1126/science.2563595. [DOI] [PubMed] [Google Scholar]
  37. Quon K. C., Marczynski G. T., Shapiro L. Cell cycle control by an essential bacterial two-component signal transduction protein. Cell. 1996 Jan 12;84(1):83–93. doi: 10.1016/s0092-8674(00)80995-2. [DOI] [PubMed] [Google Scholar]
  38. Ramakrishnan G., Newton A. FlbD of Caulobacter crescentus is a homologue of the NtrC (NRI) protein and activates sigma 54-dependent flagellar gene promoters. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2369–2373. doi: 10.1073/pnas.87.6.2369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ronson C. W., Astwood P. M., Nixon B. T., Ausubel F. M. Deduced products of C4-dicarboxylate transport regulatory genes of Rhizobium leguminosarum are homologous to nitrogen regulatory gene products. Nucleic Acids Res. 1987 Oct 12;15(19):7921–7934. doi: 10.1093/nar/15.19.7921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sanders D. A., Gillece-Castro B. L., Burlingame A. L., Koshland D. E., Jr Phosphorylation site of NtrC, a protein phosphatase whose covalent intermediate activates transcription. J Bacteriol. 1992 Aug;174(15):5117–5122. doi: 10.1128/jb.174.15.5117-5122.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sasse-Dwight S., Gralla J. D. Probing the Escherichia coli glnALG upstream activation mechanism in vivo. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8934–8938. doi: 10.1073/pnas.85.23.8934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schoenlein P. V., Gallman L. S., Winkler M. E., Ely B. Nucleotide sequence of the Caulobacter crescentus flaF and flbT genes and an analysis of codon usage in organisms with G + C-rich genomes. Gene. 1990 Sep 1;93(1):17–25. doi: 10.1016/0378-1119(90)90130-j. [DOI] [PubMed] [Google Scholar]
  43. Sommer J. M., Newton A. Sequential regulation of developmental events during polar morphogenesis in Caulobacter crescentus: assembly of pili on swarmer cells requires cell separation. J Bacteriol. 1988 Jan;170(1):409–415. doi: 10.1128/jb.170.1.409-415.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stephens C. M., Shapiro L. An unusual promoter controls cell-cycle regulation and dependence on DNA replication of the Caulobacter fliLM early flagellar operon. Mol Microbiol. 1993 Sep;9(6):1169–1179. doi: 10.1111/j.1365-2958.1993.tb01246.x. [DOI] [PubMed] [Google Scholar]
  45. Stephens C. M., Zweiger G., Shapiro L. Coordinate cell cycle control of a Caulobacter DNA methyltransferase and the flagellar genetic hierarchy. J Bacteriol. 1995 Apr;177(7):1662–1669. doi: 10.1128/jb.177.7.1662-1669.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stock J. B., Stock A. M., Mottonen J. M. Signal transduction in bacteria. Nature. 1990 Mar 29;344(6265):395–400. doi: 10.1038/344395a0. [DOI] [PubMed] [Google Scholar]
  47. Szeto W. W., Nixon B. T., Ronson C. W., Ausubel F. M. Identification and characterization of the Rhizobium meliloti ntrC gene: R. meliloti has separate regulatory pathways for activation of nitrogen fixation genes in free-living and symbiotic cells. J Bacteriol. 1987 Apr;169(4):1423–1432. doi: 10.1128/jb.169.4.1423-1432.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Terrana B., Newton A. Requirement of a cell division step for stalk formation in Caulobacter crescentus. J Bacteriol. 1976 Oct;128(1):456–462. doi: 10.1128/jb.128.1.456-462.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Van Way S. M., Newton A., Mullin A. H., Mullin D. A. Identification of the promoter and a negative regulatory element, ftr4, that is needed for cell cycle timing of fliF operon expression in Caulobacter crescentus. J Bacteriol. 1993 Jan;175(2):367–376. doi: 10.1128/jb.175.2.367-376.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Weiss D. S., Batut J., Klose K. E., Keener J., Kustu S. The phosphorylated form of the enhancer-binding protein NTRC has an ATPase activity that is essential for activation of transcription. Cell. 1991 Oct 4;67(1):155–167. doi: 10.1016/0092-8674(91)90579-n. [DOI] [PubMed] [Google Scholar]
  51. Wingrove J. A., Gober J. W. A sigma 54 transcriptional activator also functions as a pole-specific repressor in Caulobacter. Genes Dev. 1994 Aug 1;8(15):1839–1852. doi: 10.1101/gad.8.15.1839. [DOI] [PubMed] [Google Scholar]
  52. Wingrove J. A., Gober J. W. Identification of an asymmetrically localized sensor histidine kinase responsible for temporally and spatially regulated transcription. Science. 1996 Oct 25;274(5287):597–601. doi: 10.1126/science.274.5287.597. [DOI] [PubMed] [Google Scholar]
  53. Wingrove J. A., Mangan E. K., Gober J. W. Spatial and temporal phosphorylation of a transcriptional activator regulates pole-specific gene expression in Caulobacter. Genes Dev. 1993 Oct;7(10):1979–1992. doi: 10.1101/gad.7.10.1979. [DOI] [PubMed] [Google Scholar]
  54. Wozniak D. J., Ohman D. E. Pseudomonas aeruginosa AlgB, a two-component response regulator of the NtrC family, is required for algD transcription. J Bacteriol. 1991 Feb;173(4):1406–1413. doi: 10.1128/jb.173.4.1406-1413.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Yu J., Shapiro L. Early Caulobacter crescentus genes fliL and fliM are required for flagellar gene expression and normal cell division. J Bacteriol. 1992 May;174(10):3327–3338. doi: 10.1128/jb.174.10.3327-3338.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Zhuang W. Y., Shapiro L. Caulobacter FliQ and FliR membrane proteins, required for flagellar biogenesis and cell division, belong to a family of virulence factor export proteins. J Bacteriol. 1995 Jan;177(2):343–356. doi: 10.1128/jb.177.2.343-356.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Zweiger G., Marczynski G., Shapiro L. A Caulobacter DNA methyltransferase that functions only in the predivisional cell. J Mol Biol. 1994 Jan 14;235(2):472–485. doi: 10.1006/jmbi.1994.1007. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES