1887

Abstract

SUMMARY

The X174 gene product (gp) causes lysis of by inducing the host autolytic system. Experiments were carried out to ascertain which part of the 91 amino acid polypeptide carries the functional site for this process. For this purpose fusion genes were created comprising the first 51 codons of gene and unrelated sequences coding for 102 or 33 amino acids respectively. The chimeric protein of 153 amino acids consisting of the N-terminal part of gp and a fragment of -galactosidase, was neither able to lyse nor to restore -galactosidase activity by -complementation. Expression of the 84 amino acid polypeptide, however, was able to induce lysis of . It is therefore concluded that the functional lysis-inducing site of gp is located within the cloned N-terminal part of gene . In the shorter chimeric protein the sequence following the functional site was tolerated or necessary for stabilization, but in the longer chimeric protein, the C-terminal sequence disturbed the lysis-inducing conformation.

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1985-06-01
2024-05-04
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References

  1. Barrell B. G., Air G. M., Hutchison C. A. 1976; Overlapping genes in bacteriophage ϕ X174. Nature, London 264:34–41
    [Google Scholar]
  2. Bläsi U., Geisen R., Lubitz W., Henrich B., Plapp R. 1983; Localisation of the bacteriophage ϕ X174 lysis gene product in the cell envelope of Escherichia coli. In The Target of Penicillin pp 205–210 Edited by Hakenbeck R., Holtje J.-V., Labischinski H. Berlin & New York: de Gruyters;
    [Google Scholar]
  3. Fasman G. D. 1976 In Proteins; Handbook of Biochemistry and Molecular Biology vol 1: 3rd edn p 8 Edited by Fasman G. D. Cleveland: CRC Press;
    [Google Scholar]
  4. Henrich B., Lubitz W., Plapp R. 1982a; Lysis ofEscherichia coli by induction of clonedϕ X174 genes. Molecular and General Genetics 185:493–497
    [Google Scholar]
  5. Henrich B., Lubitz W., Fuchs E. 1982b; Use of benzoylated-naphthoylated DEAE-cellulose to purify and concentrate DNA eluted from agarose gels. Journal of Biochemical and Biophysical Methods 6:149–157
    [Google Scholar]
  6. Hopp T. P., Woods K. R. 1981; Prediction of protein antigenic determinants from amino acid sequences. Proceedings of the National Academy of Sciences, U,. S,. A 78:3824–3828
    [Google Scholar]
  7. Hutchison C. A. III, Sinsheimer R. L. 1966; The process of infection with bacteriophageϕ X174. Mutations in aϕ X lysis gene. Journal of Molecular Biology 18:429–447
    [Google Scholar]
  8. Lubitz W., Halfmann G., Plapp R. 1984a; Lysis ofEscherichia coli after infection withϕ X174 depends on the regulation of the cellular autolytic system. Journal of General Microbiology 130:1079–1087
    [Google Scholar]
  9. Lubitz W., Harkness R., Ishiguro E. E. 1984b; Requirement for a functional host cell autolytic enzyme system for lysis ofEscherichia coli by bacteriophageϕ X174. Journal of Bacteriology 159:385–387
    [Google Scholar]
  10. Miller J. H. 1972 Experiments in Molecular Genetics New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  11. Remaut E., Stanssens P., Fiers W. 1981; Plasmid vectors for high-efficiency expression controlled by the pL promoter of coliphage lambda. Gene 15:81–93
    [Google Scholar]
  12. Rüther U., Koenen M., Otto K., Muller-Hill B. 1981; pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucleic Acids Research 9:4087–4098
    [Google Scholar]
  13. Sanger F., Coulson A. R., Friedmann T., Air G. M., Barrell B. G., Brown N. L., Fiddes J. C., Hutchison C. A. III, Slocombe P. M., Smith M. 1978; The nucleotide sequence of bacteriophageϕ X174. Journal of Molecular Biology 125:225–246
    [Google Scholar]
  14. Sutcliffe J. G. 1979; Complete nucleotide sequence of theEscherichia coli plasmid pBR322. Cold Spring Harbor Symposia on Quantitative Biology 43:77–90
    [Google Scholar]
  15. Tessman E. S., Tessman I. 1978; The genes of the isometric phages and their functions. In The Single-stranded DNA Phages pp 9–29 Edited by Denhardt D. T., Dressier D., Ray D. S. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  16. Young K. D., Young R. 1982; Lytic action of clonedϕ X174 geneE. Journal of Virology 44:993–1002
    [Google Scholar]
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