Skip to main content
SpringerLink
Log in

Transcriptional and translational signals of the uidA gene in Escherichia coli K12

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The expression of uidA is negatively controlled by the products of the uidR and uxuR genes and is sensitive to catabolite repression. The locations of the transcriptional and translational signals of uidA were determined using lac gene fusions and S1 mapping experiments. The promoter structure of uidA resembles that of a promoter activated by cAMP receptor protein (CRP); putative control regions are located at positions-10 and-35 (relative to the transcription start site), are separated by more than 17bp and exhibit poor homology with the normally recognized consensus sequences. Moreover, 80 pb separate the promoter from the translational signals. No CRP binding site was detected in the promoter region of uidA. Two operator sites, 01 and 02, were identified: 01 has a greater affinity for the UidR repressor, whereas 02 has a greater affinity for the UxuR repressor, but the two repressor molecules are able to bind at both the 01 and 02 sites. Analysis of two operator constitutive mutations allowed the location of one of the two UidR repressor binding sites; it contains palindromic units spanning the TaqI site of the uidA control region

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aoyama T, Takanami M, Ohtsuka E, Tanimaya Y, Marumoto R, Sato H, Ikehara M (1983) Essential structure of E. coli promoter: effect of spacer length between the two consensus sequences on promoter function. Nucleic Acids Res 11:5855–5864

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1524

    Google Scholar 

  • Blanco C, Nemoz G (1987) One step purification of Escherichia coli β-glucuronidase. Biochimie 69:157–161

    Google Scholar 

  • Blanco C, Ritzenthaler P, Mata-Gilsinger M (1982) Cloning and endonuclease restriction analysis of uidA and uidR genes in Escherichia coli K-12: determination of transcription direction for the uidA gene. J Bacteriol 149:587–594

    Google Scholar 

  • Blanco C, Ritzenthaler P, Mata-Gilsinger M (1985) Nucleotide sequence of a regulatory region of the uidA gene in Escherichia coli K-12. Mol Gen Genet 199:101–105

    Google Scholar 

  • Blanco C, Ritzenthaler P, Mata-Gilsinger M (1986) Negative dominant mutations of the uidR gene in Escherichia coli: genetic proof for a cooperative regulation of uidA expression. Genetics 112:173–182

    Google Scholar 

  • Bouvier J, Patte JC, Stragier P (1984) Multiple regulatory signals in the control region of the Escherichia coli carAB operon. Proc Natl Acad Sci USA 81:4139–4143

    Google Scholar 

  • Boyer HW, Routland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472

    Google Scholar 

  • Burke JF (1984) High-sensitivity S1 mapping with single-stranded 32P DNA probes synthesized from bacteriophage M13mp template. Gene 30:63–68

    Google Scholar 

  • Busby S, Dreyfus M (1983) Segment specific mutagenesis of the regulatory region in the Escherichia coli galactose operon: isolation of mutations reducing the initiation of transcription and translation. Gene 21:121–131

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Cohen SN, Chang A, Boyer H, Helling R (1973) Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci USA 70:3240–3244

    Google Scholar 

  • De Crombrugghe B, Busby S, Buc H (1984) Activation of transcription by cyclic AMP receptor protein. In: Goldberger RF, Yamamoto KR (eds) Biological regulation and development, vol 3B. Publishing Corp, New York, pp 129–167

    Google Scholar 

  • Dente L, Cesareni G, Cortese R (1983) pEMBL: a new family of single stranded plasmids. Nucleic Acids Res 11:1645–1655

    Google Scholar 

  • Fried MG, Crothers DM (1983) CAP and RNA polymerase interactions with the lac promoter: binding stoichiometry and low range effects. Nucleic Acids Res 11:141–148

    Google Scholar 

  • Galas JD, Schmitz A (1978) DNase footprint: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res 5:3157–3170

    Google Scholar 

  • Gicquel-Sanzey B, Cossart P (1982) Homologies between different procaryotic DNA-binding regulatory proteins and between their sites of action. EMBO J 1:591–595

    Google Scholar 

  • Hawley DK, McClure WR (1983) Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255

    Google Scholar 

  • Jochimsen B, Nygaard P, Vestergaard T (1975) Location on the chromosome of Escherichia coli of genes governing purine metabolism. Mol Gen Genet 143:85–91

    Google Scholar 

  • Johnson AD, Meyer BJ, Ptashne M (1979) Interactions between DNA-bound repressors govern regulation by the phage λ repressor. Proc Natl Acad Sci USA 76:5061–5065

    Google Scholar 

  • Kolb A, Spassky A, Chapon C, Blazy B, Buc H (1983) On the different binding affinities of CRP at the lac, gal and malT promoter regions. Nucleic Acids Res 11:7833–7852

    Google Scholar 

  • Kozak M (1984) Comparison of initiation of protein synthesis in procaryotes, eukaryotes and organelles. Microbiological Rev 47:1–45

    Google Scholar 

  • Mandel M, Higa A (1970) Calcium dependent bacteriophage DNA infection. J Mol Biol 53:159–162

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Minton NP (1984) Improved plasmid vectors for the isolation of translational lac gene fusions. Gene 31:269–273

    Google Scholar 

  • Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26:101–106

    Google Scholar 

  • Novel G (1973) Le metabolisme des hexuronides chez Escherichia coli K-12. D Sci Thesis, University of Lyon, Lyon, France

    Google Scholar 

  • Novel M, Novel G (1976a) Regulation of β-glucuronidase synthesis in Escherichia coli K-12: constitutive mutants specially derepressed for uidA expression. J Bacteriol 127:406–417

    Google Scholar 

  • Novel M, Novel G (1976b) Regulation of β-glucuronidase synthesis in Escherichia coli K-12: constitutive mutations affecting uxu and uidA expression. J Bacteriol 127:418–432

    Google Scholar 

  • Radloff R, Bauer W, Vinograd J (1967) A dye-buoyant density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci USA 57:1514–1521

    Google Scholar 

  • Raibaud O, Schwartz M (1984) Positive control of transcription initiation in bacteria. Annu Rev Genet 13:319–353

    Google Scholar 

  • Ritzenthaler P, Mata-Gilsinger M, Stoeber F (1980) Construction and expression of hybrid plasmids containing Escherichia coli K-12 uxu genes. J Bacteriol 143:1116–1126

    Google Scholar 

  • Ritzenthaler P, Blanco C, Mata-Gilsinger M (1983) Interachangeability of repressors for the control of the uxu and uid operons in E. coli K-12. Mol Gen Genet 191:263–270

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Salser W, Gestland R, Bolle A (1967) In vitro synthesis of bacteriophage lysozyme. Nature 215:588–591

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sharp P, Sugden B, Sambrook J (1975) Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry 12:3055–3063

    Google Scholar 

  • Shine J, Delgarno L (1974) The 3′ terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Smitz A, Galas JD (1979) The interaction of RNA polymerase and Lac repressor with the lac control region. Nucleic Acids Res 6:111–137

    Google Scholar 

  • Stoeber F (1961) Etude des propriétés de la biosynthèse de la glucuronidase et de la glucuronide-pérméase chez Escherichia coli. D Sci Thesis, University of Paris, Paris, France

    Google Scholar 

  • Sykes RB, Nordström K (1972) Microiodometric determination of β-lactamase activity. Antimicrob Agents Chemother 1:94–99

    Google Scholar 

  • Tessman I (1968) Mutagenic treatment of double- and singlestranded DNA phages T4 and S13 with hydroxylamine. Virology 35:330–333

    Google Scholar 

  • Travers A (1980) Promoter sequence for stringent control of bacterial ribonucleic acid synthesis. J Bacteriol 141:973–976

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Microbiologie-406-I.N.S.A. de Lyon, 20 avenue Albert Eistein, F-69621, Villeurbanne Cedex, France

    Carlos Blanco

Authors
  1. Carlos Blanco
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by A. Böck

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blanco, C. Transcriptional and translational signals of the uidA gene in Escherichia coli K12. Mol Gen Genet 208, 490–498 (1987). https://doi.org/10.1007/BF00328145

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00328145

Key words

Search

Navigation