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J Bacteriol. 1985 Jul; 163(1): 94–105.
PMCID: PMC219085

Nucleotide sequence of an Escherichia coli chromosomal hemolysin.


We determined the DNA sequence of an 8,211-base-pair region encompassing the chromosomal hemolysin, molecularly cloned from an O4 serotype strain of Escherichia coli. All four hemolysin cistrons (transcriptional order, C, A, B, and D) were encoded on the same DNA strand, and their predicted molecular masses were, respectively, 19.7, 109.8, 79.9, and 54.6 kilodaltons. The identification of pSF4000-encoded polypeptides in E. coli minicells corroborated the assignment of the predicted polypeptides for hlyC, hlyA, and hlyD. However, based on the minicell results, two polypeptides appeared to be encoded on the hlyB region, one similar in size to the predicted molecular mass of 79.9 kilodaltons, and the other a smaller 46-kilodalton polypeptide. The four hemolysin gene displayed similar codon usage, which is atypical for E. coli. This reflects the low guanine-plus-cytosine content (40.2%) of the hemolysin DNA sequence and suggests the non-E. coli origin of the hemolysin determinant. In vitro-derived deletions of the hemolysin recombinant plasmid pSF4000 indicated that a region between 433 and 301 base pairs upstream of the putative start of hlyC is necessary for hemolysin synthesis. Based on the DNA sequence, a stem-loop transcription terminator-like structure (a 16-base-pair stem followed by seven uridylates) in the mRNA was predicted distal to the C-terminal end of hlyA. A model for the general transcriptional organization of the E. coli hemolysin determinant is presented.

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  • Berger H, Hacker J, Juarez A, Hughes C, Goebel W. Cloning of the chromosomal determinants encoding hemolysin production and mannose-resistant hemagglutination in Escherichia coli. J Bacteriol. 1982 Dec;152(3):1241–1247. [PMC free article] [PubMed]
  • Cavalieri SJ, Snyder IS. Effect of Escherichia coli alpha-hemolysin on human peripheral leukocyte viability in vitro. Infect Immun. 1982 May;36(2):455–461. [PMC free article] [PubMed]
  • Cavalieri SJ, Snyder IS. Effect of Escherichia coli alpha-hemolysin on human peripheral leukocyte function in vitro. Infect Immun. 1982 Sep;37(3):966–974. [PMC free article] [PubMed]
  • Cavalieri SJ, Snyder IS. Cytotoxic activity of partially purified Escherichia coli alpha haemolysin. J Med Microbiol. 1982 Feb;15(1):11–21. [PubMed]
  • Chou PY, Fasman GD. Prediction of protein conformation. Biochemistry. 1974 Jan 15;13(2):222–245. [PubMed]
  • Felmlee T, Pellett S, Lee EY, Welch RA. Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J Bacteriol. 1985 Jul;163(1):88–93. [PMC free article] [PubMed]
  • Gadeberg OV, Orskov I. In vitro cytotoxic effect of alpha-hemolytic Escherichia coli on human blood granulocytes. Infect Immun. 1984 Jul;45(1):255–260. [PMC free article] [PubMed]
  • Gill RE, Heffron F, Falkow S. Identification of the protein encoded by the transposable element Tn3 which is required for its transposition. Nature. 1979 Dec 20;282(5741):797–801. [PubMed]
  • Goebel W, Hedgpeth J. Cloning and functional characterization of the plasmid-encoded hemolysin determinant of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1290–1298. [PMC free article] [PubMed]
  • Gribskov M, Devereux J, Burgess RR. The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):539–549. [PMC free article] [PubMed]
  • Grosjean H, Fiers W. Preferential codon usage in prokaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene. 1982 Jun;18(3):199–209. [PubMed]
  • Hacker J, Hughes C, Hof H, Goebel W. Cloned hemolysin genes from Escherichia coli that cause urinary tract infection determine different levels of toxicity in mice. Infect Immun. 1983 Oct;42(1):57–63. [PMC free article] [PubMed]
  • Härtlein M, Schiessl S, Wagner W, Rdest U, Kreft J, Goebel W. Transport of hemolysin by Escherichia coli. J Cell Biochem. 1983;22(2):87–97. [PubMed]
  • Hawley DK, McClure WR. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. [PMC free article] [PubMed]
  • Hull SI, Hull RA, Minshew BH, Falkow S. Genetics of hemolysin of Escherichia coli. J Bacteriol. 1982 Aug;151(2):1006–1012. [PMC free article] [PubMed]
  • Ikemura T. Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes: a proposal for a synonymous codon choice that is optimal for the E. coli translational system. J Mol Biol. 1981 Sep 25;151(3):389–409. [PubMed]
  • Juarez A, Goebel W. Chromosomal mutation that affects excretion of hemolysin in Escherichia coli. J Bacteriol. 1984 Sep;159(3):1083–1085. [PMC free article] [PubMed]
  • Juarez A, Hughes C, Vogel M, Goebel W. Expression and regulation of the plasmid-encoded hemolysin determinant of Escherichia coli. Mol Gen Genet. 1984;197(2):196–203. [PubMed]
  • Knapp S, Hacker J, Then I, Müller D, Goebel W. Multiple copies of hemolysin genes and associated sequences in the chromosomes of uropathogenic Escherichia coli strains. J Bacteriol. 1984 Sep;159(3):1027–1033. [PMC free article] [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Levy SB. R factor proteins synthesized in Escherichia coli minicells: incorporation studies with different R factors and detection of deoxyribonucleic acid-binding proteins. J Bacteriol. 1974 Dec;120(3):1451–1463. [PMC free article] [PubMed]
  • Mackman N, Holland IB. Secretion of a 107 K dalton polypeptide into the medium from a haemolytic E. coli K12 strain. Mol Gen Genet. 1984;193(2):312–315. [PubMed]
  • MARMUR J, DOTY P. Thermal renaturation of deoxyribonucleic acids. J Mol Biol. 1961 Oct;3:585–594. [PubMed]
  • Messing J, Crea R, Seeburg PH. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. [PMC free article] [PubMed]
  • Michaelis S, Beckwith J. Mechanism of incorporation of cell envelope proteins in Escherichia coli. Annu Rev Microbiol. 1982;36:435–465. [PubMed]
  • Noegel A, Rdest U, Goebel W. Determination of the functions of hemolytic plasmid pHly152 of Escherichia coli. J Bacteriol. 1981 Jan;145(1):233–247. [PMC free article] [PubMed]
  • Noegel A, Rdest U, Springer W, Goebel W. Plasmid cistrons controlling synthesis and excretion of the exotoxin alpha-haemolysin of Escherichia coli. Mol Gen Genet. 1979 Oct 1;175(3):343–350. [PubMed]
  • Perlman D, Halvorson HO. A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol. 1983 Jun 25;167(2):391–409. [PubMed]
  • POLLOCK MR, RICHMOND MH. Low cyst(e)ine content of bacterial extracellular proteins: its possible physiological significance. Nature. 1962 May 5;194:446–449. [PubMed]
  • Pustell J, Kafatos FC. A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):643–655. [PMC free article] [PubMed]
  • Rosenberg M, Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. [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]
  • Shine J, Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. [PMC free article] [PubMed]
  • Silhavy TJ, Benson SA, Emr SD. Mechanisms of protein localization. Microbiol Rev. 1983 Sep;47(3):313–344. [PMC free article] [PubMed]
  • Stark JM, Shuster CW. Analysis of hemolytic determinants of plasmid pHly185 by Tn5 mutagenesis. J Bacteriol. 1982 Nov;152(2):963–967. [PMC free article] [PubMed]
  • Tinoco I, Jr, Borer PN, Dengler B, Levin MD, Uhlenbeck OC, Crothers DM, Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. [PubMed]
  • Vieira J, Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. [PubMed]
  • von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. [PubMed]
  • Wagner W, Vogel M, Goebel W. Transport of hemolysin across the outer membrane of Escherichia coli requires two functions. J Bacteriol. 1983 Apr;154(1):200–210. [PMC free article] [PubMed]
  • Welch RA, Dellinger EP, Minshew B, Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. [PubMed]
  • Welch RA, Falkow S. Characterization of Escherichia coli hemolysins conferring quantitative differences in virulence. Infect Immun. 1984 Jan;43(1):156–160. [PMC free article] [PubMed]
  • Welch RA, Hull R, Falkow S. Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect Immun. 1983 Oct;42(1):178–186. [PMC free article] [PubMed]
  • Yamamoto T, Tamura T, Yokota T. Primary structure of heat-labile enterotoxin produced by Escherichia coli pathogenic for humans. J Biol Chem. 1984 Apr 25;259(8):5037–5044. [PubMed]

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