• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Dec 1996; 178(23): 6975–6982.
PMCID: PMC178601

Identification of sequences necessary for transcription in vitro from the Chlamydia trachomatis rRNA P1 promoter.

Abstract

Chlamydia trachomatis RNA polymerase was partially purified by heparin-agarose chromatography and used in conjunction with a plasmid-borne G-less cassette template to characterize the C. trachomatis rRNA P1 promoter in vitro. Stepwise mutational analysis revealed that sequences in the -10, -25, and -35 regions are necessary for promoter activity, but no sequence upstream of position -40 is required. Partially purified C. trachomatis RNA polymerase and purified Escherichia coli holoenzyme exhibited some differences in promoter specificity.

Full Text

The Full Text of this article is available as a PDF (456K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allen JE, Stephens RS. Identification by sequence analysis of two-site posttranslational processing of the cysteine-rich outer membrane protein 2 of Chlamydia trachomatis serovar L2. J Bacteriol. 1989 Jan;171(1):285–291. [PMC free article] [PubMed]
  • Brickman TJ, Barry CE, 3rd, Hackstadt T. Molecular cloning and expression of hctB encoding a strain-variant chlamydial histone-like protein with DNA-binding activity. J Bacteriol. 1993 Jul;175(14):4274–4281. [PMC free article] [PubMed]
  • Chamberlin M, Kingston R, Gilman M, Wiggs J, deVera A. Isolation of bacterial and bacteriophage RNA polymerases and their use in synthesis of RNA in vitro. Methods Enzymol. 1983;101:540–568. [PubMed]
  • Dombroski AJ, Walter WA, Record MT, Jr, Siegele DA, Gross CA. Polypeptides containing highly conserved regions of transcription initiation factor sigma 70 exhibit specificity of binding to promoter DNA. Cell. 1992 Aug 7;70(3):501–512. [PubMed]
  • Douglas AL, Saxena NK, Hatch TP. Enhancement of in vitro transcription by addition of cloned, overexpressed major sigma factor of Chlamydia psittaci 6BC. J Bacteriol. 1994 May;176(10):3033–3039. [PMC free article] [PubMed]
  • Elliott T, Geiduschek EP. Defining a bacteriophage T4 late promoter: absence of a "-35" region. Cell. 1984 Jan;36(1):211–219. [PubMed]
  • Engel JN, Ganem D. A polymerase chain reaction-based approach to cloning sigma factors from eubacteria and its application to the isolation of a sigma-70 homolog from Chlamydia trachomatis. J Bacteriol. 1990 May;172(5):2447–2455. [PMC free article] [PubMed]
  • Engel JN, Pollack J, Malik F, Ganem D. Cloning and characterization of RNA polymerase core subunits of Chlamydia trachomatis by using the polymerase chain reaction. J Bacteriol. 1990 Oct;172(10):5732–5741. [PMC free article] [PubMed]
  • Engel JN, Pollack J, Perara E, Ganem D. Heat shock response of murine Chlamydia trachomatis. J Bacteriol. 1990 Dec;172(12):6959–6972. [PMC free article] [PubMed]
  • Engel JN, Ganem D. Chlamydial rRNA operons: gene organization and identification of putative tandem promoters. J Bacteriol. 1987 Dec;169(12):5678–5685. [PMC free article] [PubMed]
  • Everett KD, Hatch TP. Sequence analysis and lipid modification of the cysteine-rich envelope proteins of Chlamydia psittaci 6BC. J Bacteriol. 1991 Jun;173(12):3821–3830. [PMC free article] [PubMed]
  • Fahr MJ, Douglas AL, Xia W, Hatch TP. Characterization of late gene promoters of Chlamydia trachomatis. J Bacteriol. 1995 Aug;177(15):4252–4260. [PMC free article] [PubMed]
  • Fahr MJ, Sriprakash KS, Hatch TP. Convergent and overlapping transcripts of the Chlamydia trachomatis 7.5-kb plasmid. Plasmid. 1992 Nov;28(3):247–257. [PubMed]
  • Gaal T, Ross W, Blatter EE, Tang H, Jia X, Krishnan VV, Assa-Munt N, Ebright RH, Gourse RL. DNA-binding determinants of the alpha subunit of RNA polymerase: novel DNA-binding domain architecture. Genes Dev. 1996 Jan 1;10(1):16–26. [PubMed]
  • Gadeau AP, Mouches C, Bove JM. Probable insensitivity of mollicutes to rifampin and characterization of spiroplasmal DNA-dependent RNA polymerase. J Bacteriol. 1986 Jun;166(3):824–828. [PMC free article] [PubMed]
  • Gu L, Wenman WM, Remacha M, Meuser R, Coffin J, Kaul R. Chlamydia trachomatis RNA polymerase alpha subunit: sequence and structural analysis. J Bacteriol. 1995 May;177(9):2594–2601. [PMC free article] [PubMed]
  • Hackstadt T, Baehr W, Ying Y. Chlamydia trachomatis developmentally regulated protein is homologous to eukaryotic histone H1. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3937–3941. [PMC free article] [PubMed]
  • Helmann JD. Compilation and analysis of Bacillus subtilis sigma A-dependent promoter sequences: evidence for extended contact between RNA polymerase and upstream promoter DNA. Nucleic Acids Res. 1995 Jul 11;23(13):2351–2360. [PMC free article] [PubMed]
  • Koehler JE, Burgess RR, Thompson NE, Stephens RS. 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]
  • Lambden PR, Everson JS, Ward ME, Clarke IN. Sulfur-rich proteins of Chlamydia trachomatis: developmentally regulated transcription of polycistronic mRNA from tandem promoters. Gene. 1990 Mar 1;87(1):105–112. [PubMed]
  • Mathews SA, Douglas A, Sriprakash KS, Hatch TP. In vitro transcription in Chlamydia psittaci and Chlamydia trachomatis. Mol Microbiol. 1993 Mar;7(6):937–946. [PubMed]
  • Mathews SA, Sriprakash KS. The RNA polymerase of Chlamydia trachomatis has a flexible sequence requirement at the -10 and -35 boxes of its promoters. J Bacteriol. 1994 Jun;176(12):3785–3789. [PMC free article] [PubMed]
  • Mathews SA, Sriprakash KS. A strand-specific endonucleolytic activity with DNA site preference for cleavage in Chlamydia trachomatis. J Bacteriol. 1994 Aug;176(15):4774–4778. [PMC free article] [PubMed]
  • Moulder JW. Interaction of chlamydiae and host cells in vitro. Microbiol Rev. 1991 Mar;55(1):143–190. [PMC free article] [PubMed]
  • Parvin JD, Sharp PA. Identification of novel factors which bind specifically to the core promoter of the immunoglobulin heavy chain gene. J Biol Chem. 1991 Dec 5;266(34):22878–22886. [PubMed]
  • Perara E, Ganem D, Engel JN. A developmentally regulated chlamydial gene with apparent homology to eukaryotic histone H1. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2125–2129. [PMC free article] [PubMed]
  • Reznikoff WS, Siegele DA, Cowing DW, Gross CA. The regulation of transcription initiation in bacteria. Annu Rev Genet. 1985;19:355–387. [PubMed]
  • Ross W, Gosink KK, Salomon J, Igarashi K, Zou C, Ishihama A, Severinov K, Gourse RL. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Sawadogo M, Roeder RG. Factors involved in specific transcription by human RNA polymerase II: analysis by a rapid and quantitative in vitro assay. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4394–4398. [PMC free article] [PubMed]
  • Schachter J. The intracellular life of Chlamydia. Curr Top Microbiol Immunol. 1988;138:109–139. [PubMed]
  • Tam JE, Davis CH, Wyrick PB. Expression of recombinant DNA introduced into Chlamydia trachomatis by electroporation. Can J Microbiol. 1994 Jul;40(7):583–591. [PubMed]
  • Tan M, Wong B, Engel JN. Transcriptional organization and regulation of the dnaK and groE operons of Chlamydia trachomatis. J Bacteriol. 1996 Dec;178(23):6983–6990. [PMC free article] [PubMed]
  • Upender M, Raj L, Weir M. Megaprimer method for in vitro mutagenesis using parallel templates. Biotechniques. 1995 Jan;18(1):29–32. [PubMed]
  • Wichlan DG, Hatch TP. Identification of an early-stage gene of Chlamydia psittaci 6BC. J Bacteriol. 1993 May;175(10):2936–2942. [PMC free article] [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...