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GENETICS AND MOLECULAR BIOLOGY

Identification of a Conserved N-Terminal Sequence Involved in Transmembrane Signal Transduction in EnvZ

Jill Waukau, Steven Forst
Jill Waukau
Department of Biological Sciences, University of Wisconsin, Milwaukee, Wisconsin 53201
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Steven Forst
Department of Biological Sciences, University of Wisconsin, Milwaukee, Wisconsin 53201
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DOI: 10.1128/JB.181.17.5534-5538.1999
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  • Fig. 1.
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    Fig. 1.

    Comparison of amino acid sequences of the Met1-to-Asp244 regions of various EnvZ proteins. Invariant residues in the N-terminal region (Met1 to Ile179) are indicated by stars beneath the sequence. The I box is enclosed in a box. The TM1 and TM2 domains and the sequence containing the H243 site of phosphorylation (H box) are underlined. Amino acid substitutions of mutants isolated in this study are indicated by arrowheads above the sequence.

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    Fig. 2.

    Outer membrane protein analysis. AT142pcnBcells containing various envZ alleles were grown on MacConkey medium, and outer membrane proteins were prepared and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as described previously (5). Lane 1, pJW49 (wild type); lane 2, pJW32P; lane 3, pJW35P; lane 4, pJW43P; lane 5, pJW48E. C, OmpC; F, OmpF; A, OmpA.

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  • Table 1.

    Bacterial strains and plasmids

    Strain or plasmidRelevant genotypeSource or reference
    Strains
     S. flexneriWild typeATCC 120022
     Enterobacter cloacaeWild typeATCC 23355
     Y. enterocoliticaWild typeLab strain
     P. vulgarisWild typeATCC 13315
     E. coli
      AT142F−envZ::Kan lacU169 araD139 rpsL relA thiA flbB12
      AT142pcnBpcnB::Tn10of AT14212
      WH57ompF-lacZof AT1427
      LEO544pcnB::Tn10 of WH5712
    Plasmids
     pMRL25ompR-envZ in pBR32212
     pKS2pMRL25 with envZH243N19
     pJW49pMRL25 with aSalI site at bp 5–10 of envZThis study
     pJW32PpJW49 with envZ L32PThis study
     pJW35ApMRL25 with envZ L35AThis study
     pJW35PpJW49 with envZ L35PThis study
     pJW43ApMRL25 with envZ L43AThis study
     pJW43PpJW49 with envZ L43PThis study
     pJW48EpJW49 with envZ K48EThis study
  • Table 2.

    Amino acid sequence identity relative to theE. coli protein

    Bacterium% Identity for:
    EnvZaI box
    S. flexneri99.6100
    Enterobacter cloacae98.4100
    Y. enterocolitica92.2100
    P. vulgaris45.8100
    V. choleraeb35.785
    • ↵a Met1 to Ile179.

    • ↵b Taken from reference 23.

  • Table 3.

    Porin protein production on MacConkey medium

    StrainRelative % of porin protein produceda
    OmpFOmpC
    Wild type1434
    L32P440
    L35P243
    L43P240
    K48E151
    • ↵a The values represent the relative amounts of OmpF and OmpC, expressed as a percentage of the total absorbance of OmpA, OmpF, and OmpC, as determined by scanning densitometry of the outer membrane proteins.

  • Table 4.

    Porin protein production in nutrient broth medium

    StrainRelative % of porin protein produced ina:
    NBNB + sucrose
    OmpFOmpCOmpFOmpC
    Wild type5118753
    L35P1948850
    L43P1750655
    L35A5420360
    L43A1858266
    • ↵a The values represent the relative amounts of OmpF and OmpC as described in the footnote in Table 3. NB, nutrient broth.

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Identification of a Conserved N-Terminal Sequence Involved in Transmembrane Signal Transduction in EnvZ
Jill Waukau, Steven Forst
Journal of Bacteriology Sep 1999, 181 (17) 5534-5538; DOI: 10.1128/JB.181.17.5534-5538.1999

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Identification of a Conserved N-Terminal Sequence Involved in Transmembrane Signal Transduction in EnvZ
Jill Waukau, Steven Forst
Journal of Bacteriology Sep 1999, 181 (17) 5534-5538; DOI: 10.1128/JB.181.17.5534-5538.1999
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KEYWORDS

Bacterial Outer Membrane Proteins
Conserved Sequence
Escherichia coli Proteins
Multienzyme Complexes
Phosphoprotein Phosphatases
signal transduction

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