Summary
The haemolysin (hly) determinant of the plasmid pHly152 contains an IS2 element at 469 bp upstream of the hlyC gene. The sequence at the other (right-hand) end (RS) also shows multiple hybridization with the plasmid pHly152 and the chromosome of some Escherichia coli strains but the nucleotide sequence of this region does not reveal the typical properties of an IS element. Similar arrangements in the regions flanking the hly determinant are also found on various Hly plasmids from uropathogenic E. coli strains. Chromosomal hly determinants lack both flanking sequences (IS2 and RS) in the immediate vicinity pf the hly genes. The sequences immediately upstream of the hlyC gene have been determined from several chromosomal hly determinants and compared with the corresponding sequence of the hly determinant of the plasmid pHly152. We show that these sequences, which contain one promoter (left promoter, phly L) in all hly determinants tested, vary considerably although common sequence elements can still be identified. In contrast, only relatively few nucleotide exchanges have been detected in the adjacent structural hlyC genes. The A+T content of the 200 bp sequence upstream of hlyC is very high (72 mol% A+T) but even the structural hly genes show a considerably higher A+T content (about 60 mol%) than the E. coli chromosome on average (50 mol% A+T) suggesting that the hly determinant may not have originated in E. coli.
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References
Beaucage SL, Caruthers MH (1981) Deoxynucleoside phosphoramidites: a new class of key intermediates for deoxypolynucleotide synthesis. Tetrahedron Lett 22:1859–1862
Berger H, Hacker J, Juarez A, Hughes C, Goebel W (1982) Cloning of the chromosomal determinants encoding hemolysin production and mannose resistant hemagglutination in Escherichia coli. J Bacteriol 152:1241–1247
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acid Res 7:1513–1523
Collins J, Brüning HJ (1978) Plasmids useable as gene cloning vectors in an in vitro packaging by coliphage lambda: “cosmids”. Gene 4:85–107
De la Cruz F, Müller D, Ortiz JM, Goebel W (1980) A hemolysin determinant common to Escherichia coli Hly plasmids of different uncompatibility groups. J Bacteriol 143:825–833
Ghosal D, Sommer H, Saedler H (1979) Nucleotide sequence of the transposable DNA element IS2. Nucleic Acids Res 6:1111–1112
Goebel W, Hedgpeth J (1982) Cloning and functional characterization of the entire plasmid-encoded hemolysin determinant of Escherichia coli. J Bacteriol 151:1290–1298
Goebel W, Royer-Pokora B, Lindenmair W, Bujard H (1974) Plasmids controlling synthesis of hemolysis in Escherichia coli: molecular properties. J Bacteriol 118:964–973
Grinstedt J, Bennett PM, Higginson S, Richmond MH (1978) Regional preference of insertion of TN501 and Tn802 into RP1 and its derivatives. Mol Gen Genet 166:313–320
Hacker J, Knapp S, Goebel W (1983a) Spontaneous deletions and flanking regions of the chromosomally inherited hemolysin determinant of an Escherichia coli 06 strain. J Bacteriol 154:1145–1152
Hacker J, Hughes C, Hof H, Goebel W (1983b) Cloned hemolysin genes from Escherichia coli that cause urinary tract infection determine different levels of toxicity in mice. Infect Immunol 42:57–63
Hacker J, Hughes C (1985) Genetics of Escherichia coli hemolysin. Curr Top Microbiol Immunol, in press
Hu N, Messing J (1982) The making of strand-specific M13 probes. Gene 17:271–277
Juarez A, Hughes C, Vogel M, Goebel W (1984) Expression and regulation of the plasmid-encoded hemolysin determinant of Escherichia coli. Mol Gen Genet 197:196–203
Knapp S, Hacker J, Then I, Müller D, Goebel W (1984) Multiple copies of hemolysin genes and associated sequences in the chromosome of uropathogenic Escherichia coli strains. J Bacteriol 159:1027–1033
Lederberg EM, Cohen SN (1974) Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol 119:1072–1074
Mackman N, Holland B (1984) Functional characterization of a cloned haemolysin determinant from E. coli of human origin, encoding information for the secretion of a 107 K polypeptide. Mol Gen Genet 196:129–134
Michel G (1984) Feinkartierung und Sequenzierung eines Endes der Plasmidkodierten Hämolysindeterminante von Escherichia coli. Diplom Thesis, University Würzburg
Minshew BH, Jorgensen J, Counts GW, Falkow S (1978) Association of hemolysin production, hemagglutination of human erythrocytes, and virulence for chicken embryos of extraintestinal Escherichia coli isolates. Infect Immunol 20:50–54
Müller D, Hughes C, Goebel W (1983) Relationship between plasmid and chromosomal hemolysin determinants of Escherichia coli. J Bacteriol 153:846–851
Noegel A, Rdest U, Goebel W (1981) Determination of the functions of hemolytic plasmid pHly152 of Escherichia coli. J Bacteriol 145:233–247
Rigby PW, Dieckman JM, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Smith HW, Halls S (1967) The transmissible nature of the genetic factor in Escherichia coli that controls haemolysin production. J Gen Microbiol 47:153–161
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stark JM, Shuster CW (1982) Analysis of hemolytic determinants of plasmid pHly152 by Tn5 mutagenesis. J Bacteriol 152:963–967
Tietze E, Tschäpe H (1983) Plasmid pattern analysis of natural bacterial isolates and its epidemiological implication. J Hyg Camb 90:475–488
Wagner W, Vogel M, Goebel W (1983) Transport of hemolysin across the outer membrane of Escherichia coli requires two functions. J Bacteriol 154:200–210
Welch RA, Falkow S (1984) Characterization of Escherichia coli hemolysins conferring quantitative differences in virulence. Infect Immunol 43:156–160
Welch RA, Dellinger EP, Minshew B, Falkow S (1981) Hemolysin contributes to virulence of extraintestinal Escherichia coli infections. Nature 294:665–667
Welch RA, Hull R, Falkow S (1983) Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect Immunol 42:178–186
Zabala JC, De la Cruz F, Ortiz JM (1982) Several copies of the same insertion sequence are present on alpha-hemolytic plasmids belonging to four different inompatibility groups. J Bacteriol 151:472–476
Zabala JC, Garcia-Lobo JM, Diaz-Aroca E, De la Cruz F, Ortiz M (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
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Communicated by E. Bautz
Dedicated to Prof. F. Lingens on the occasion of his 60th anniversary
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Knapp, S., Then, I., Wels, W. et al. Analysis of the flanking regions from different haemolysin determinants of Escherichia coli . Molec. Gen. Genet. 200, 385–392 (1985). https://doi.org/10.1007/BF00425721
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DOI: https://doi.org/10.1007/BF00425721