Skip to main content
SpringerLink
Log in

Comparison of the haemolysin secretion protein HlyB from Proteus vulgaris and Escherichia coli; site-directed mutagenesis causing impairment of export function

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

Summary

The hlyB secretion genes of Proteus vulgaris and Escherichia coli showed 81% nucleotide homology and similar E. coli-atypical codon usage. The deduced protein sequences differed in 54 of 707 residues and shared a previously unreported sequence which corresponds to the ATP-binding motif characteristic of protein kinases. The motif was also conserved in the HlyB of Morganella morganii. Of 4 oligonucleotide-directed substitutions introduced into the putative E. coli HlyB motif, 2 non-conservative changes caused radical reductions in the export of active haemolysin protein.

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

References

  • Booher R, Beach DH (1986) Site-specific mutagenesis of cdc 2+, a cell cycle control gene of the fission yeast Schizosaccharomyces pombe. Mol Cell Biol 6:3523–3530

    Google Scholar 

  • Chen L, Tai PC (1987) Evidence for the involvement of ATP in co-translational protein translocation. Nature 328:164–166

    Google Scholar 

  • Citron BA, Donelson JE (1984) Sequence of the Saccharomyces GAL region and its transcription in vivo. J Bacteriol 158:269–278

    Google Scholar 

  • Debouck C, Riccio D, Schumperli D, McKenney K, Jeffers J, Hughes C, Rosenberg M, Heusterspreute M, Brunel F, Davidson J (1985) Structure of the galactokinase gene of Escherichia coli, the last (?) gene of the gal operon. Nucleic Acids Res 13:1841–1853

    Google Scholar 

  • Eilers M, Oppliger W, Schatz G (1987) Both ATP and an energized inner membrane are required to import a purified precursor protein into mitochondria. EMBO J 6:1073–1077

    Google Scholar 

  • Felmlee T, Pellett S, Lee E-Y, Welch R (1985a) Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J Bacteriol 163:88–93

    Google Scholar 

  • Felmlee T, Pellett S, Welch R (1985b) Nucleotide sequence of an Escherichia coli chromosomal hemolysin. J Bacteriol 163:94–105

    Google Scholar 

  • Gerlach JH, Endicott JA, Juranka PF, Henderson G, Sarangi F, Deuchars KL, Ling V (1986) Homology between P-glycoprotein and a bacterial haemolysin transport protein suggests a model for multidrug resistance. Nature 324:485–489

    Google Scholar 

  • Haertlein M, Schießl S, Wagner W, Rdest U, Kreft J, Goebel W (1983) Transport of hemolysin by Escherichia coli. J Cell Biochem 22:87–97

    Google Scholar 

  • Hashimoto E, Takio E, Krebs EG (1982) Amino acid sequence at a ATP-binding site of cGMP-dependent protein kinase. J Biol Chem 257:727–733

    Google Scholar 

  • Hess J, Wels W, Vogel M, Goebel W (1986) Nucleotide sequence of a plasmid-encoded hemolysin determinant and its comparison with a corresponding chromosomal hemolysin sequence. FEMS Microbiol Lett 34:1–11

    Google Scholar 

  • Higgins CF, Hiles ID, Salmond GC, Gill DR, Downie JA, Evans IJ, Holland IB, Gray L, Buckel S, Bell AW, Hermodson MA (1986) A family of related ATP-binding subunits coupled to many distinct biological processes in bacteria Nature 323:448–450

    Google Scholar 

  • Kitamura N, Kitamura A, Toyoshima K, Hirayama Y, Yoshida M (1982) Avian sarcoma virus Y73 genome sequence and structural similarity of its transforming gene product to that of Rous sarcoma virus. Nature 297:205–208

    Google Scholar 

  • Koronakis V, Cross M, Koronakis E, Senior B, Hughes C (1987) The secreted hemolysins of Proteus mirabilis, Proteus vulgaris and Morganella morganii are genetically related to each other and to the alpha-hemolysin of Escherichia coli. J Bacteriol 169:1509–1515

    Google Scholar 

  • Kramer W, Drutsa V, Jansen HW, Kramer B, Pflugfelder M, Fritz HJ (1984) The gapped duplex DNA approach to oligonucleotide-directed mutation construction. Nucleic Acids Res 12:9441–9456

    Google Scholar 

  • Mackman N, Nicaud J-M, Gray L, Holland IB (1986) Secretion of hemolysin by Escherichia coli. Curr Top Microbiol Immunol 125:159–181

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–580

    Google Scholar 

  • Perara, E, Rothman RE, Lingappa VR (1986) Uncoupling translocation from translation: implications for transport of proteins across membrane. Science 232:348–352

    Google Scholar 

  • Privalsky ML, Ralston R, Bishop JM (1984) The membrane glycoprotein encoded by the retroviral oncogene v-erb-B is structurally related to tyrosine-specific protein kinases. Proc Natl Acad Sci USA 81:704–707

    Google Scholar 

  • Reddy EP, Smith MJ, Srinivasan A (1983) Nucleotide sequence of Abelson murine leukemia virus genome; structural similarity of its transforming gene product to other one gene products with tyrosine-specific kinase activity. Proc Natl Acad Sci USA 80:3623–3627

    Google Scholar 

  • Rosenthal A, Rhee L, Yadegari Y, Paro R, Ullric A, Goeddel DV (1987) Structure and nucleotide sequence of a Drosophila melanogaster protein kinase C gene. EMBO J 6:433–441

    Google Scholar 

  • Rothman JE, Kornberg RD, (1986) An unfolding story of protein translocation. Nature 322:209–210

    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 

  • Schwartz D, Tizard R, Gilbert W (1983) Nucleotide sequence of Rous sarcoma virus. Cell 32:853–869

    Google Scholar 

  • Shibuya M, Hanafusa H (1982) Nucleotide sequence of Fujinami sarcoma virus; evolutionary relationship of its transforming gene with transforming genes of other sarcoma viruses. Cell 30:787–795

    Google Scholar 

  • Shoji S, Parmelee DC, Wade R, Kuma S, Ericsson L, Walsh K, Neurath H, Long G, Demaille J, Fischer E, Titani K (1981) Complete amino acid sequence of the catalytic subunit of bovine cardiac muscle cyclic AMP-dependent protein kinase. Proc Natl Acad Sci USA 78:848–851

    Google Scholar 

  • Van Beveren C, Galleshaw J, Jonas V, Berns V, Doolittle RF, Donoghue D, Verma I (1981) Nucleotide sequence and formation of the transforming gene of a mouse sarcoma virus. Nature 289:258–262

    Google Scholar 

  • Wagner W, Vogel M, Goebel W (1983) Transport of hemolysin across the outer membrane of Escherchia coli requires two functions. J Bacteriol 154:200–210

    Google Scholar 

  • Zabala JC, Garcia-Lobo J, Diaz-Aroca E, Cruz dela F, Ortiz JM (1984) Escherichia coli alpha-hemolysin synthesis and export genes are flanked by a direct repetition of IS91-like elements. Mol Gen Genet 197:90–97

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, Cambridge University, Tennis Court Road, CB2 1QP, Cambridge, England

    Vassilis Koronakis, Eva Koronakis & Colin Hughes

Authors
  1. Vassilis Koronakis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eva Koronakis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Colin Hughes
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by J.W. Lengeler

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koronakis, V., Koronakis, E. & Hughes, C. Comparison of the haemolysin secretion protein HlyB from Proteus vulgaris and Escherichia coli; site-directed mutagenesis causing impairment of export function. Mol Gen Genet 213, 551–555 (1988). https://doi.org/10.1007/BF00339631

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00339631

Key words

Search

Navigation