Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Apr;173(7):2301–2310. doi: 10.1128/jb.173.7.2301-2310.1991

Negative regulatory loci coupling flagellin synthesis to flagellar assembly in Salmonella typhimurium.

K L Gillen 1, K T Hughes 1
PMCID: PMC207783  PMID: 1848842

Abstract

The complex regulation of flagellin gene expression in Salmonella typhimurium was characterized in vivo by using lac transcriptional fusions to the two flagellin structural genes (fliC [H1] and fljB [H2]). Phase variation was measured as the rate of switching of flagellin gene expression. Switching frequencies varied from 1/500 per cell per generation to 1/10,000 per cell per generation depending on the particular insertion and the direction of switching. There is a 4- to 20-fold bias in favor of switching from the fljB(On) to the fljB(Off) orientation. Random Tn10dTc insertions were isolated which failed to express flagellin. While most of these insertions mapped to loci known to be required for flagellin expression, several new loci were identified. The presence of functional copies of all of the genes responsible for complete flagellar assembly, except the hook-associated proteins (flgK, flgL, and fliD gene products), were required for expression of the fliC or fljB flagellin genes. Two novel loci involved in negative regulation of fliC and fljB in fla mutant backgrounds were identified. One of these loci, designated the flgR locus, mapped to the flg operon at 23 min on the Salmonella linkage map. An flgR insertion mutation resulted in relief of repression of the fliC and fljB genes in all fla mutant backgrounds except for mutants in the positive regulatory loci (flhC, flhD, and fliA genes).

Full text

PDF
2301

Selected References

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

  1. Adler J., Templeton B. The effect of environmental conditions on the motility of Escherichia coli. J Gen Microbiol. 1967 Feb;46(2):175–184. doi: 10.1099/00221287-46-2-175. [DOI] [PubMed] [Google Scholar]
  2. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bochner B. R., Savageau M. A. Generalized indicator plate for genetic, metabolic, and taxonomic studies with microorganisms. Appl Environ Microbiol. 1977 Feb;33(2):434–444. doi: 10.1128/aem.33.2.434-444.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bruist M. F., Simon M. I. Phase variation and the Hin protein: in vivo activity measurements, protein overproduction, and purification. J Bacteriol. 1984 Jul;159(1):71–79. doi: 10.1128/jb.159.1.71-79.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chan R. K., Botstein D., Watanabe T., Ogata Y. Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high-frequency-transducing lysate. Virology. 1972 Dec;50(3):883–898. doi: 10.1016/0042-6822(72)90442-4. [DOI] [PubMed] [Google Scholar]
  8. Eismann E., von Wilcken-Bergmann B., Müller-Hill B. Specific destruction of the second lac operator decreases repression of the lac operon in Escherichia coli fivefold. J Mol Biol. 1987 Jun 20;195(4):949–952. doi: 10.1016/0022-2836(87)90499-2. [DOI] [PubMed] [Google Scholar]
  9. Flashner Y., Gralla J. D. Dual mechanism of repression at a distance in the lac operon. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8968–8972. doi: 10.1073/pnas.85.23.8968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glasgow A. C., Bruist M. F., Simon M. I. DNA-binding properties of the Hin recombinase. J Biol Chem. 1989 Jun 15;264(17):10072–10082. [PubMed] [Google Scholar]
  11. Haber R., Adhya S. Interaction of spatially separated protein-DNA complexes for control of gene expression: operator conversions. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9683–9687. doi: 10.1073/pnas.85.24.9683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hamilton E. P., Lee N. Three binding sites for AraC protein are required for autoregulation of araC in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1749–1753. doi: 10.1073/pnas.85.6.1749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Heichman K. A., Johnson R. C. The Hin invertasome: protein-mediated joining of distant recombination sites at the enhancer. Science. 1990 Aug 3;249(4968):511–517. doi: 10.1126/science.2166334. [DOI] [PubMed] [Google Scholar]
  14. Hughes K. T., Roth J. R. Conditionally transposition-defective derivative of Mu d1(Amp Lac). J Bacteriol. 1984 Jul;159(1):130–137. doi: 10.1128/jb.159.1.130-137.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hughes K. T., Roth J. R. Transitory cis complementation: a method for providing transposition functions to defective transposons. Genetics. 1988 May;119(1):9–12. doi: 10.1093/genetics/119.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hughes K. T., Youderian P., Simon M. I. Phase variation in Salmonella: analysis of Hin recombinase and hix recombination site interaction in vivo. Genes Dev. 1988 Aug;2(8):937–948. doi: 10.1101/gad.2.8.937. [DOI] [PubMed] [Google Scholar]
  17. Huo L., Martin K. J., Schleif R. Alternative DNA loops regulate the arabinose operon in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5444–5448. doi: 10.1073/pnas.85.15.5444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Iino T. Genetics and chemistry of bacterial flagella. Bacteriol Rev. 1969 Dec;33(4):454–475. doi: 10.1128/br.33.4.454-475.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Iino T. Genetics of structure and function of bacterial flagella. Annu Rev Genet. 1977;11:161–182. doi: 10.1146/annurev.ge.11.120177.001113. [DOI] [PubMed] [Google Scholar]
  20. Iino T., Komeda Y., Kutsukake K., Macnab R. M., Matsumura P., Parkinson J. S., Simon M. I., Yamaguchi S. New unified nomenclature for the flagellar genes of Escherichia coli and Salmonella typhimurium. Microbiol Rev. 1988 Dec;52(4):533–535. doi: 10.1128/mr.52.4.533-535.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Johnson R. C., Simon M. I. Hin-mediated site-specific recombination requires two 26 bp recombination sites and a 60 bp recombinational enhancer. Cell. 1985 Jul;41(3):781–791. doi: 10.1016/s0092-8674(85)80059-3. [DOI] [PubMed] [Google Scholar]
  22. Jones C. J., Macnab R. M. Flagellar assembly in Salmonella typhimurium: analysis with temperature-sensitive mutants. J Bacteriol. 1990 Mar;172(3):1327–1339. doi: 10.1128/jb.172.3.1327-1339.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Komeda Y. Fusions of flagellar operons to lactose genes on a mu lac bacteriophage. J Bacteriol. 1982 Apr;150(1):16–26. doi: 10.1128/jb.150.1.16-26.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Komeda Y., Iino T. Regulation of expression of the flagellin gene (hag) in Escherichia coli K-12: analysis of hag-lac gene fusions. J Bacteriol. 1979 Sep;139(3):721–729. doi: 10.1128/jb.139.3.721-729.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Komeda Y., Suzuki H., Ishidsu J. I., Iino T. The role of cAMP in flagellation of Salmonella typhimurium. Mol Gen Genet. 1976 Dec 31;142(4):289–298. doi: 10.1007/BF00271253. [DOI] [PubMed] [Google Scholar]
  26. Komeda Y. Transcriptional control of flagellar genes in Escherichia coli K-12. J Bacteriol. 1986 Dec;168(3):1315–1318. doi: 10.1128/jb.168.3.1315-1318.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kutsukake K., Iino T. Refined genetic analysis of the region II che mutants in Salmonella typhimurium. Mol Gen Genet. 1985;199(3):406–409. doi: 10.1007/BF00330750. [DOI] [PubMed] [Google Scholar]
  28. Kutsukake K., Ohya Y., Iino T. Transcriptional analysis of the flagellar regulon of Salmonella typhimurium. J Bacteriol. 1990 Feb;172(2):741–747. doi: 10.1128/jb.172.2.741-747.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kutsukake K., Ohya Y., Yamaguchi S., Iino T. Operon structure of flagellar genes in Salmonella typhimurium. Mol Gen Genet. 1988 Sep;214(1):11–15. doi: 10.1007/BF00340172. [DOI] [PubMed] [Google Scholar]
  30. Lederberg J. Detection of Fermentative Variants with Tetrazolium. J Bacteriol. 1948 Nov;56(5):695–695. doi: 10.1128/jb.56.5.695-695.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Maloy S. R., Roth J. R. Regulation of proline utilization in Salmonella typhimurium: characterization of put::Mu d(Ap, lac) operon fusions. J Bacteriol. 1983 May;154(2):561–568. doi: 10.1128/jb.154.2.561-568.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ohnishi K., Kutsukake K., Suzuki H., Iino T. Gene fliA encodes an alternative sigma factor specific for flagellar operons in Salmonella typhimurium. Mol Gen Genet. 1990 Apr;221(2):139–147. doi: 10.1007/BF00261713. [DOI] [PubMed] [Google Scholar]
  34. Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, Edition VI. Microbiol Rev. 1983 Sep;47(3):410–453. doi: 10.1128/mr.47.3.410-453.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, edition VII. Microbiol Rev. 1988 Dec;52(4):485–532. doi: 10.1128/mr.52.4.485-532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Silverman M., Simon M. Characterization of Escherichia coli flagellar mutants that are insensitive to catabolite repression. J Bacteriol. 1974 Dec;120(3):1196–1203. doi: 10.1128/jb.120.3.1196-1203.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Silverman M., Simon M. Genetic analysis of flagellar mutants in Escherichia coli. J Bacteriol. 1973 Jan;113(1):105–113. doi: 10.1128/jb.113.1.105-113.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Silverman M., Simon M. Phase variation: genetic analysis of switching mutants. Cell. 1980 Apr;19(4):845–854. doi: 10.1016/0092-8674(80)90075-6. [DOI] [PubMed] [Google Scholar]
  39. Silverman M., Zieg J., Simon M. Flagellar-phase variation: isolation of the rh1 gene. J Bacteriol. 1979 Jan;137(1):517–523. doi: 10.1128/jb.137.1.517-523.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Smith H. O. Defective phage formation by lysogens of integration deficient phage P22 mutants. Virology. 1968 Feb;34(2):203–223. doi: 10.1016/0042-6822(68)90231-6. [DOI] [PubMed] [Google Scholar]
  41. Somers J. M., Kay W. W. Genetic fine structure of the tricarboxylate transport (tct) locus of Salmonella typhimurium. Mol Gen Genet. 1983;190(1):20–26. doi: 10.1007/BF00330319. [DOI] [PubMed] [Google Scholar]
  42. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  43. Valentin-Hansen P., Albrechtsen B., Løve Larsen J. E. DNA-protein recognition: demonstration of three genetically separated operator elements that are required for repression of the Escherichia coli deoCABD promoters by the DeoR repressor. EMBO J. 1986 Aug;5(8):2015–2021. doi: 10.1002/j.1460-2075.1986.tb04458.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  45. Winkelman J. W., Clark D. P. Proton suicide: general method for direct selection of sugar transport- and fermentation-defective mutants. J Bacteriol. 1984 Nov;160(2):687–690. doi: 10.1128/jb.160.2.687-690.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yamaguchi S., Fujita H., Ishihara A., Aizawa S., Macnab R. M. Subdivision of flagellar genes of Salmonella typhimurium into regions responsible for assembly, rotation, and switching. J Bacteriol. 1986 Apr;166(1):187–193. doi: 10.1128/jb.166.1.187-193.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yamaguchi S., Fujita H., Taira T., Kutsukake K., Homma M., Iino T. Genetic analysis of three additional fla genes in Salmonella typhimurium. J Gen Microbiol. 1984 Dec;130(12):3339–3342. doi: 10.1099/00221287-130-12-3339. [DOI] [PubMed] [Google Scholar]
  48. Yokota T., Gots J. S. Requirement of adenosine 3', 5'-cyclic phosphate for flagella formation in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1970 Aug;103(2):513–516. doi: 10.1128/jb.103.2.513-516.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Youderian P., Sugiono P., Brewer K. L., Higgins N. P., Elliott T. Packaging specific segments of the Salmonella chromosome with locked-in Mud-P22 prophages. Genetics. 1988 Apr;118(4):581–592. doi: 10.1093/genetics/118.4.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Zieg J., Hilmen M., Simon M. Regulation of gene expression by site-specific inversion. Cell. 1978 Sep;15(1):237–244. doi: 10.1016/0092-8674(78)90098-3. [DOI] [PubMed] [Google Scholar]
  51. Zieg J., Silverman M., Hilmen M., Simon M. Recombinational switch for gene expression. Science. 1977 Apr 8;196(4286):170–172. doi: 10.1126/science.322276. [DOI] [PubMed] [Google Scholar]
  52. Zieg J., Simon M. Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4196–4200. doi: 10.1073/pnas.77.7.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES