1887

Microbiology Society logo

Volume 134, Issue 10

Chromosomal Mutations That Increase the Production of a Plasmid-encoded Haemolysin in Free

Abstract

Transposon mutagenesis was used to isolate two mutants which express very large amounts of haemolysin when carrying the multicopy plasmid pANN202-312. strain Hha-2 was isolated by Mu1 mutagenesis, and strain Hha-3 by Tn mutagenesis. The transposon insertion was chromosomal in both mutants and could be demonstrated to be unrelated to the haemolytic region of the plasmid. The substantial increase in both extracellular and intracellular haemolysin production was dependent upon plasmid copy number and was drastically reduced when either mutant carried the low-copy-number haemolytic plasmid pHly152. In both mutants, the marked increase in extracellular production was dependent upon the specific haemolysin transport genes, and The lack of either gene function resulted in no external haemolysin production. SDS-PAGE analysis showed no change in the pattern of outer-membrane proteins of the mutants, although changes (differing between the two mutants) were seen in their periplasmic proteins. The mutations of both strains (termed and ) were mapped at minute 10·5 of the chromosome. No relation to any known gene affecting gene regulation in could be found.

  • Received:
  • Revised:
  • Published Online:
© Society for General Microbiology, 1988
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-10-2779
1988-10-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/10/mic-134-10-2779.html?itemId=/content/journal/micro/10.1099/00221287-134-10-2779&mimeType=html&fmt=ahah

References

  1. Ames G.F.-L., Spudich E.N., Nikaido H. 1973; Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations.. Journal of Bacteriology 117:406–416
     [Google Scholar]
  2. Berger H., Hacker J., Juárez , Hughes C., Goebel W. 1982; Cloning of the chromosomal determinants encoding hemolysin production and mannose-resistant hemagglutination in Escherichia coli.. Journal of Bacteriology 152:1241–1247
     [Google Scholar]
  3. Bhakdi S., Mackman N., Nicaud J.M., Holland I.B. 1986; Escherichia coli hemolysin may damage target cell membranes by generating transmembrane pores.. Infection and Immunity 52:63–69
     [Google Scholar]
  4. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Research 7:1513–1522
     [Google Scholar]
  5. Bishai W.R., Reppuoli R., Murphy J.R. 1987; High-level expression of a proteolytically sensitive diphtheria toxin fragment in Escherichia coli.. Journal of Bacteriology 169:5140–5151
     [Google Scholar]
  6. Casadaban M.J., Cohen S. 1979; Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences.. Proceedings of the National Academy of Sciences of the United States of America 76:4530–4533
     [Google Scholar]
  7. Delacruz F., Müller D., Ortiz J.M., Goebel W. 1980; Hemolysis determinant common to Escherichia coli Hly plasmids of different incompatibility groups.. Journal of Bacteriology 143:825–833
     [Google Scholar]
  8. Felmlee T., Pellett S., Lee E.-Y., Welch R.A. 1985; Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide.. Journal of Bacteriology 163:88–93
     [Google Scholar]
  9. Filip C., Fletcher G., Wulff J.C., Earhart C.F. 1973; Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium lauryl sarcosinate.. Journal of Bacteriology 115:717–722
     [Google Scholar]
  10. Goebel W., Hedgpeth J. 1982; Cloning and functional characterization of the plasmid encoded hemolysin determinant of Escherichia coli.. Journal of Bacteriology 151:1290–1298
     [Google Scholar]
  11. González-Carrero M.I., Zabala J.C., Delacruz F., Ortiz J.M. 1985; Purification of a-hemolysin from an over-producing E. coli strain.. Molecular and General Genetics 199:106–110
     [Google Scholar]
  12. Hartlein M., Schiebl S., Wagner W., Rdest U., Kreft J., Goebel W. 1983; Transport of hemolysin by Escherichia coli.. Journal of Cell Biochemistry 22:87–97
     [Google Scholar]
  13. Juárez , Goebel W. 1984; Chromosomal mutation that affects the excretion of hemolysin in Escherichia coli.. Journal of Bacteriology 159:1083–1085
     [Google Scholar]
  14. Juárez A., Hartlein M., Goebel W. 1984a; Study of regulation and transport of hemolysin by using fusion of the β-galactosidase gene (lacZ) to hemolysin genes.. Journal of Bacteriology 160:161–168
     [Google Scholar]
  15. Juárez , Hughes C., Vogel M., Goebel W. 1984b; Expression and regulation of the plasmid encoded hemolysin determinant of Escherichia coli.. Molecular and General Genetics 197:196–203
     [Google Scholar]
  16. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature, London 227:680–685
     [Google Scholar]
  17. L6Fdahl S., Guss B., Uhlen B., Philipson M., Lindberg M. 1983; Gene for staphylococcal protein A.. Proceedings of the National Academy of Sciences of the United States of America 80:697–701
     [Google Scholar]
  18. Mackman N., Mackman N., Holland I. B. 1984; Secretion of a 107 kilodalton polypeptide into the medium from a haemolytic Escherichia coli K12 strain.. Molecular and General Genetics 196:129–134
     [Google Scholar]
  19. Mackman N., Nicaud J.M., Gray L., Holland I.B. 1985a; Genetical and functional organization of the Escherichia coli haemolysin determinant 2001.. Molecular and General Genetics 201:282–288
     [Google Scholar]
  20. Mackman N., Nicaud J.M., Gray L., Holland I.B. 1985b; Identification of polypeptides required for the export of haemolysin 2001 from E. coli.. Molecular and General Genetics 201:529–536
     [Google Scholar]
  21. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  22. Maurizi M.R., Trisler P., Gottesman S. 1985; Insertional mutagenesis of the Ion gene in Escherichia coli: Ion is dispensable.. Journal of Bacteriology 164:1124–1135
     [Google Scholar]
  23. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  24. Müller D., Hughes C., Goebel W. 1983; Relationship between plasmid and chromosomal hemolysin determinants of Escherichia coli.. Journal of Bacteriology 153:846–851
     [Google Scholar]
  25. Mulligan J.T., Margolin W., Heilig J., Walker G.C. 1982; Mutations affecting regulation of methionine biosynthetic genes isolated by use of met-lac fusions.. Journal of Bacteriology 151:609–619
     [Google Scholar]
  26. Nicaud J.M., Mackman NGRAY, Holland I.B. 1985; Characterization of HlyC and mechanism of activation and secretion of haemolysin from Escherichia coli 2001.. FEBS Letters 187:339–344
     [Google Scholar]
  27. Nieto J.M., Tomás J., Juárez A. 1987; Secretion of an Aeromonas hydrophila aerolysin by a mutant strain of Escherichia coli.. FEMS Microbiology letters 48:413–417
     [Google Scholar]
  28. Noegel A., Rdest U., Goebel W. 1981; Determination of the functions of hemolytic plasmid pHlyl52 of Escherichia coli.. Journal of Bacteriology 145:233–247
     [Google Scholar]
  29. O’Hanley P., Low D., Romero I., Lark D., Vosti K., Falkow S., Schoolnik G. 1985; Gal-Gal binding and hemolysin phenotypes and genotypes associated with uropathogenic Escherichia coli.. New England Journal of Medicine 313:414–420
     [Google Scholar]
  30. Shaw K.J., Berg C.M. 1979; Escherichia coli auxotrophs induced by insertion of the transposable element Tn5.. Genetics 92:741–747
     [Google Scholar]
  31. Talmagde K., Stahl S., Gilbert W. 1980; Eukaryotic signal sequence transports insulin antigen in Escherichia coli.. Proceedings of the National Academy of Sciences of the United States of America 77:3369–3373
     [Google Scholar]
  32. Trisler P., Gottesman S. 1985; Ion transcriptional regulation of genes necessary for capsular polysaccharide synthesis in Escherichia coli K12.. Journal of Bacteriology 160:184–191
     [Google Scholar]
  33. Tsukagoshi N., Ihara N., Yamagata H., Udaka S. 1984; Cloning and expession of a thermophilic a-amylase gene from Bacillus stearothermophilus in Escherichia coli.. Molecular and General Genetics 193:58–63
     [Google Scholar]
  34. Vogel M., Hess J., Then I., Juárez A., Goebel W. 1988; Characterization of a sequence (Hly R) which enhances synthesis and secretion of haemolysin in Escherichia coli.. Molecular and General Genetics 212:76–84
     [Google Scholar]
  35. Wagner W., Vogel M., Goebel W. 1983; Transport of hemolysin across the outer membrane of Escherichia coli requires two functions.. Journal of Bacteriology 154:200–210
     [Google Scholar]
  36. Wanner B.L. 1986; Novel regulatory mutants of the phosphate regulon in Escherichia coli K12.. Journal of Molecular Biology 191:39–58
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-10-2779
Loading
Chromosomal Mutations That Increase the Production of a Plasmid-encoded Haemolysin in Escherichia coli
Microbiology 134, 2779 (1988); https://doi.org/10.1099/00221287-134-10-2779
/content/journal/micro/10.1099/00221287-134-10-2779
/content/journal/micro/10.1099/00221287-134-10-2779
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error