Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Aug;177(16):4645–4651. doi: 10.1128/jb.177.16.4645-4651.1995

Identification of the minimum regulatory region of a Myxococcus xanthus A-signal-dependent developmental gene.

P Gulati 1, D Xu 1, H B Kaplan 1
PMCID: PMC177228  PMID: 7642490

Abstract

Developmental expression of the Myxococcus xanthus gene 4521 requires extracellular A-signal. This signal is generated in response to nutrient limitation and functions in cell density sensing. To identify the upstream limit of the minimum region required in vivo for A-signal-dependent 4521 expression, a 5' deletion analysis of the 4521 regulatory region was performed. A new vector, pHBK280, was designed to facilitate this analysis. This vector creates tandem copies of the 4521 gene in the M. xanthus chromosome, such that the regulatory region to be tested is upstream of a single copy of the lacZ reporter gene. The 5' deletion analysis revealed that at most, 146 bp of DNA upstream of the transcription start site (TSS) was required for full developmental expression of 4521. Basal expression levels were observed with constructions containing 90 bp of DNA upstream of the TSS. In vitro gel retardation assays revealed that DNA fragments with 5' ends of 146 and 125 bp upstream of the TSS and a common 3' end of +24 bp were retarded in their mobility after incubation with all of the M. xanthus developmental crude cell extracts tested. In contrast, a fragment starting at 90 bp upstream of the TSS and ending at +24 bp was not retarded in its mobility after incubation with the same cell extracts. These in vivo and in vitro data suggest that cis-acting elements located between 146 and 90 bp upstream of the TSS serve as binding sites for one or more trans-acting regulatory factors required for 4521 developmental expression.

Full Text

The Full Text of this article is available as a PDF (369.4 KB).

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  3. Gill R. E., Cull M. G., Fly S. Genetic identification and cloning of a gene required for developmental cell interactions in Myxococcus xanthus. J Bacteriol. 1988 Nov;170(11):5279–5288. doi: 10.1128/jb.170.11.5279-5288.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gober J. W., Shapiro L. Integration host factor is required for the activation of developmentally regulated genes in Caulobacter. Genes Dev. 1990 Sep;4(9):1494–1504. doi: 10.1101/gad.4.9.1494. [DOI] [PubMed] [Google Scholar]
  5. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  6. Kaplan H. B., Kuspa A., Kaiser D. Suppressors that permit A-signal-independent developmental gene expression in Myxococcus xanthus. J Bacteriol. 1991 Feb;173(4):1460–1470. doi: 10.1128/jb.173.4.1460-1470.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kashefi K., Hartzell P. L. Genetic suppression and phenotypic masking of a Myxococcus xanthus frzF- defect. Mol Microbiol. 1995 Feb;15(3):483–494. doi: 10.1111/j.1365-2958.1995.tb02262.x. [DOI] [PubMed] [Google Scholar]
  8. Keseler I. M., Kaiser D. An early A-signal-dependent gene in Myxococcus xanthus has a sigma 54-like promoter. J Bacteriol. 1995 Aug;177(16):4638–4644. doi: 10.1128/jb.177.16.4638-4644.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kim S. K., Kaiser D., Kuspa A. Control of cell density and pattern by intercellular signaling in Myxococcus development. Annu Rev Microbiol. 1992;46:117–139. doi: 10.1146/annurev.mi.46.100192.001001. [DOI] [PubMed] [Google Scholar]
  10. Kroos L., Kuspa A., Kaiser D. A global analysis of developmentally regulated genes in Myxococcus xanthus. Dev Biol. 1986 Sep;117(1):252–266. doi: 10.1016/0012-1606(86)90368-4. [DOI] [PubMed] [Google Scholar]
  11. Kuspa A., Kroos L., Kaiser D. Intercellular signaling is required for developmental gene expression in Myxococcus xanthus. Dev Biol. 1986 Sep;117(1):267–276. doi: 10.1016/0012-1606(86)90369-6. [DOI] [PubMed] [Google Scholar]
  12. Kuspa A., Plamann L., Kaiser D. A-signalling and the cell density requirement for Myxococcus xanthus development. J Bacteriol. 1992 Nov;174(22):7360–7369. doi: 10.1128/jb.174.22.7360-7369.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuspa A., Plamann L., Kaiser D. Identification of heat-stable A-factor from Myxococcus xanthus. J Bacteriol. 1992 May;174(10):3319–3326. doi: 10.1128/jb.174.10.3319-3326.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kustu S., Santero E., Keener J., Popham D., Weiss D. Expression of sigma 54 (ntrA)-dependent genes is probably united by a common mechanism. Microbiol Rev. 1989 Sep;53(3):367–376. doi: 10.1128/mr.53.3.367-376.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Plamann L., Davis J. M., Cantwell B., Mayor J. Evidence that asgB encodes a DNA-binding protein essential for growth and development of Myxococcus xanthus. J Bacteriol. 1994 Apr;176(7):2013–2020. doi: 10.1128/jb.176.7.2013-2020.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Plamann L., Kuspa A., Kaiser D. Proteins that rescue A-signal-defective mutants of Myxococcus xanthus. J Bacteriol. 1992 May;174(10):3311–3318. doi: 10.1128/jb.174.10.3311-3318.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Plamann L., Li Y., Cantwell B., Mayor J. The Myxococcus xanthus asgA gene encodes a novel signal transduction protein required for multicellular development. J Bacteriol. 1995 Apr;177(8):2014–2020. doi: 10.1128/jb.177.8.2014-2020.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scanlan D. J., Bloye S. A., Mann N. H., Hodgson D. A., Carr N. G. Construction of lacZ promoter probe vectors for use in Synechococcus: application to the identification of CO2-regulated promoters. Gene. 1990 May 31;90(1):43–49. doi: 10.1016/0378-1119(90)90437-v. [DOI] [PubMed] [Google Scholar]
  21. Shimkets L. J. Social and developmental biology of the myxobacteria. Microbiol Rev. 1990 Dec;54(4):473–501. doi: 10.1128/mr.54.4.473-501.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wireman J. W., Dworkin M. Morphogenesis and developmental interactions in myxobacteria. Science. 1975 Aug 15;189(4202):516–523. doi: 10.1126/science.806967. [DOI] [PubMed] [Google Scholar]
  23. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES