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. 1994 Jun;176(12):3785–3789. doi: 10.1128/jb.176.12.3785-3789.1994

The RNA polymerase of Chlamydia trachomatis has a flexible sequence requirement at the -10 and -35 boxes of its promoters.

S A Mathews 1, K S Sriprakash 1
PMCID: PMC205568  PMID: 8206857

Abstract

Mutated variants of the predicted promoter of the countertranscript of the Chlamydia trachomatis plasmid were tested by in vitro transcription with chlamydial extract. A 3-bp deletion within the -10 region of the putative promoter caused the RNA polymerase to initiate transcription 3 bases downstream. Many single mutations in the -10 and -35 regions did not alter promoter function. However, some multiple mutations in both hexamers rendered the promoter inefficient or ineffective. Taken together, these results indicate that (i) the sequence requirement for chlamydial promoters differs from that for Escherichia coli and (ii) chlamydial RNA polymerase can tolerate considerably more variation at the -10 and -35 regions. These results are paradoxical considering the homology between C. trachomatis sigma 66 and E. coli sigma 70.

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Selected References

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

  1. Amann E., Brosius J., Ptashne M. Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene. 1983 Nov;25(2-3):167–178. doi: 10.1016/0378-1119(83)90222-6. [DOI] [PubMed] [Google Scholar]
  2. Birkelund S., Lundemose A. G., Christiansen G. The 75-kilodalton cytoplasmic Chlamydia trachomatis L2 polypeptide is a DnaK-like protein. Infect Immun. 1990 Jul;58(7):2098–2104. doi: 10.1128/iai.58.7.2098-2104.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chiang L. W., Kovari I., Howe M. M. Mutagenic oligonucleotide-directed PCR amplification (Mod-PCR): an efficient method for generating random base substitution mutations in a DNA sequence element. PCR Methods Appl. 1993 Feb;2(3):210–217. doi: 10.1101/gr.2.3.210. [DOI] [PubMed] [Google Scholar]
  4. Comanducci M., Cevenini R., Moroni A., Giuliani M. M., Ricci S., Scarlato V., Ratti G. Expression of a plasmid gene of Chlamydia trachomatis encoding a novel 28 kDa antigen. J Gen Microbiol. 1993 May;139(5):1083–1092. doi: 10.1099/00221287-139-5-1083. [DOI] [PubMed] [Google Scholar]
  5. Crenshaw R. W., Fahr M. J., Wichlan D. G., Hatch T. P. Developmental cycle-specific host-free RNA synthesis in Chlamydia spp. Infect Immun. 1990 Oct;58(10):3194–3201. doi: 10.1128/iai.58.10.3194-3201.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Engel J. N., Ganem D. Chlamydial rRNA operons: gene organization and identification of putative tandem promoters. J Bacteriol. 1987 Dec;169(12):5678–5685. doi: 10.1128/jb.169.12.5678-5685.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Erickson J. W., Gross C. A. Identification of the sigma E subunit of Escherichia coli RNA polymerase: a second alternate sigma factor involved in high-temperature gene expression. Genes Dev. 1989 Sep;3(9):1462–1471. doi: 10.1101/gad.3.9.1462. [DOI] [PubMed] [Google Scholar]
  8. Fahr M. J., Sriprakash K. S., Hatch T. P. Convergent and overlapping transcripts of the Chlamydia trachomatis 7.5-kb plasmid. Plasmid. 1992 Nov;28(3):247–257. doi: 10.1016/0147-619x(92)90056-g. [DOI] [PubMed] [Google Scholar]
  9. Gao J. G., Gussin G. N. RNA polymerases from Pseudomonas aeruginosa and Pseudomonas syringae respond to Escherichia coli activator proteins. J Bacteriol. 1991 Jan;173(1):394–397. doi: 10.1128/jb.173.1.394-397.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gardella T., Moyle H., Susskind M. M. A mutant Escherichia coli sigma 70 subunit of RNA polymerase with altered promoter specificity. J Mol Biol. 1989 Apr 20;206(4):579–590. doi: 10.1016/0022-2836(89)90567-6. [DOI] [PubMed] [Google Scholar]
  11. Graña D., Gardella T., Susskind M. M. The effects of mutations in the ant promoter of phage P22 depend on context. Genetics. 1988 Oct;120(2):319–327. doi: 10.1093/genetics/120.2.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Han C. Y., Crawford I. P., Harwood C. S. Up-promoter mutations in the trpBA operon of Pseudomonas aeruginosa. J Bacteriol. 1991 Jun;173(12):3756–3762. doi: 10.1128/jb.173.12.3756-3762.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hatch T. P., Vance D. W., Jr, Al-Hossainy E. Identification of a major envelope protein in Chlamydia spp. J Bacteriol. 1981 Apr;146(1):426–429. doi: 10.1128/jb.146.1.426-429.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kaul R., Gray G. J., Koehncke N. R., Gu L. J. Cloning and sequence analysis of the Chlamydia trachomatis spc ribosomal protein gene cluster. J Bacteriol. 1992 Feb;174(4):1205–1212. doi: 10.1128/jb.174.4.1205-1212.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koehler J. E., Burgess R. R., Thompson N. E., Stephens R. S. Chlamydia trachomatis RNA polymerase major sigma subunit. Sequence and structural comparison of conserved and unique regions with Escherichia coli sigma 70 and Bacillus subtilis sigma 43. J Biol Chem. 1990 Aug 5;265(22):13206–13214. [PubMed] [Google Scholar]
  17. Lambden P. R., Everson J. S., Ward M. E., Clarke I. N. Sulfur-rich proteins of Chlamydia trachomatis: developmentally regulated transcription of polycistronic mRNA from tandem promoters. Gene. 1990 Mar 1;87(1):105–112. doi: 10.1016/0378-1119(90)90500-q. [DOI] [PubMed] [Google Scholar]
  18. Lonetto M., Gribskov M., Gross C. A. The sigma 70 family: sequence conservation and evolutionary relationships. J Bacteriol. 1992 Jun;174(12):3843–3849. doi: 10.1128/jb.174.12.3843-3849.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mathews S. A., Douglas A., Sriprakash K. S., Hatch T. P. In vitro transcription in Chlamydia psittaci and Chlamydia trachomatis. Mol Microbiol. 1993 Mar;7(6):937–946. doi: 10.1111/j.1365-2958.1993.tb01185.x. [DOI] [PubMed] [Google Scholar]
  20. Moulder J. W. Interaction of chlamydiae and host cells in vitro. Microbiol Rev. 1991 Mar;55(1):143–190. doi: 10.1128/mr.55.1.143-190.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pearce B. J., Fahr M. J., Hatch T. P., Sriprakash K. S. A chlamydial plasmid is differentially transcribed during the life cycle of Chlamydia trachomatis. Plasmid. 1991 Sep;26(2):116–122. doi: 10.1016/0147-619x(91)90051-w. [DOI] [PubMed] [Google Scholar]
  22. Ricci S., Cevenini R., Cosco E., Comanducci M., Ratti G., Scarlato V. Transcriptional analysis of the Chlamydia trachomatis plasmid pCT identifies temporally regulated transcripts, anti-sense RNA and sigma 70-selected promoters. Mol Gen Genet. 1993 Mar;237(3):318–326. doi: 10.1007/BF00279434. [DOI] [PubMed] [Google Scholar]
  23. Sardinia L. M., Engel J. N., Ganem D. Chlamydial gene encoding a 70-kilodalton antigen in Escherichia coli: analysis of expression signals and identification of the gene product. J Bacteriol. 1989 Jan;171(1):335–341. doi: 10.1128/jb.171.1.335-341.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sriprakash K. S., Pearce B. J. Mapping of transcripts encoded by the plasmid in Chlamydia trachomatis. FEMS Microbiol Lett. 1990 Sep 15;59(3):299–303. doi: 10.1016/0378-1097(90)90237-k. [DOI] [PubMed] [Google Scholar]
  25. Tanaka K., Takahashi H. Cloning and analysis of the gene (rpoDA) for the principal sigma factor of Pseudomonas aeruginosa. Biochim Biophys Acta. 1991 May 2;1089(1):113–119. doi: 10.1016/0167-4781(91)90092-z. [DOI] [PubMed] [Google Scholar]
  26. Yuan Y., Zhang Y. X., Manning D. S., Caldwell H. D. Multiple tandem promoters of the major outer membrane protein gene (omp1) of Chlamydia psittaci. Infect Immun. 1990 Sep;58(9):2850–2855. doi: 10.1128/iai.58.9.2850-2855.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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