Summary
The hemolytic activity of Escherichia coli and Proteus vulgaris is determined by common contiguous genes encoding synthesis (hly C, hly A) and specific secretion (hly B, hly D) of active hemolysin. Nevertheless, the hlyC-proximal DNA sequences directing production of the homologous hemolysins by the recombinant DNAs P. vulgaris pVU763-709 and E. coli pANN202-312 showed no extensive homology. Primer extension and S1 nuclease protection were used to define in the two sequences the 5′ termini of hly transcripts synthesized in vivo and thus to infer the active hly promoters sequences. The E. coli hly C upstream region contained three separate promotors directing in vivo hly transcription, while the corresponding transcription of the P. vulgaris hly operon originated from a single distinct promotor, the-35 and-10 sequences of which formed part of an inverted repeat sequence. Elevated hemolytic activity caused by upstream Tn5 insertions in pVU763-709 resulted from increased transcription from this promotor.
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References
Berg DE, Weiss A, Crossland L (1980) Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol 142:439–446
de Crombugghe B, Pastan I (1980) Cyclic AMP, the cyclic AMP receptor protein and the dual control of the galactose operon. In: Miller J, Reznikoff W (eds) The Operon. Cold Spring Harbor Laboratory Press, New York, pp 303–324
Felmlee T, Pellett S, Welch RA (1985a) Nucleotide sequence of an Escherichia coli chromosomal hemolysin. J Bacteriol 163:94–105
Felmlee T, Pellett S, Lee E-Y, Welch RA (1985b) Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J Bacteriol 163:88–93
Gerlach JH, Endicott J, Juranka P, Henderson G, Sarangi F, Deuchars K, Ling V (1986) Homology between P-glycoprotein and a bacterial hemolysin protein suggests a model for multidrug resistance. Nature 324:485–489
Gonzalez-Carrero MI, Zabala JC, De la Cruz F, Ortiz JM (1985) Purification of α-hemolysin from an overproducing E. coli strain. Mol Gen Genet 199:106–110
Gunsales RP, Yanofsky C (1980) Nucleotide sequence and expression of Escherichia coli trp R, the structural gene for the trp aporepressor. Proc Natl Acad Sci USA 12:7117–7121
Hacker J, Hughes C (1985) Genetics of Escherichia coli hemolysin. Curr Top Microbiol Immunol 118:139–162
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
Juarez A, Haertlein M, Goebel W (1984a) Fusion of the β-galactosidase gene (lacZ) to hemolysin genes to study regulation and secretion of Escherichia coli hemolysin. J Bacteriol 160:161–168
Juarez A, Hughes C, Vogel M, Goebel w (1984b) Expression and regulation of the plasmid-encoded hemolysin determinant of Escherichia coli. Mol Gen Genet 197:196–203
Kleckner N (1981) Transposable elements. Annu Rev Genet 15:341–404
Knapp S, Hacker J, Then I, Muller D, Goebel W (1984) Multiple copies of hemolysin genes and associated sequences in the chromosomes of uropathogenic Escherichia coli strains. J Bacteriol 159:1027–1033
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
Mackman N, Nicaud J-M, Gray L, Holland IB (1985) Genetical and functional organisation of the Escherichia coli haemolysin determinant 2001. Mol Gen Genet 201:282–288
Mackman N, Nicaud J-M, Gray L, Holland IB (1986) Secretion of haemolysin by Escherichia coli. Curr Top Microbiol Immunol 125:159–181
Mackman N, Baker K, Gray L, Haigh R, Nicaud J-M, Holland IB (1987) Release of a chimeric protein into the medium from Escherichia coli using the C-terminal secretion signal of haemolysin. EMBO J 6:2835–2842
Maxam AM, Gibert W (1980) Sequencing end-labelled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Muller D, Hughes C, Goebel W (1983) Relationship between plasmid and chromosomal hemolysin determinants of E. coli. J Bacteriol 153:846–851
Nicaud J-M, Mackman N, Gray L, Holland IB (1985a) Characterization of HlyC and mechanism of activation and secretion of hemolysin from E. coli 2001. FEBS Lett 187:339–334
Nicaud J-M, Mackman N, Gray L, Holland IB (1985b) Regulation of haemolysin synthesis in E. coli determined by hly genes of human origin. Mol Gen Genet 199:111–116
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Staden R (1982) An interactive graphics program for comparing and aligning nucleic acid and amino acid sequence. Nucleic Acids Res 10:2951–2961
Wagner W, Vogel M, Goebel W (1983) Transport of haemolysin across the outer membrane of Escherichia coli requires two functions. J Bacteriol 154:200–210
Weaver RF, Weissman C (1979) Mapping of RNA by a modification of the Berk-Sharp procedure: the 5′ termini of 15 S beta-globin mRNA precursor and mature 10S beta-globin mRNA have identical map coordinates. Nucleic Acids Res 7:1175–1193
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Communicated by J.W. Lengeler
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Koronakis, V., Hughes, C. Identification of the promotors directing in vivo expression of hemolysin genes in Proteus vulgaris and Escherichia coli . Molec. Gen. Genet. 213, 99–104 (1988). https://doi.org/10.1007/BF00333404
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DOI: https://doi.org/10.1007/BF00333404