Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1980 Apr;34(1):67–72. doi: 10.1128/jvi.34.1.67-72.1980

Temperature-sensitive mutants of foot-and-mouth disease virus with altered structural polypeptides. II. Comparison of recombination and biochemical maps.

A M King, W R Slade, J W Newman, D McCahon
PMCID: PMC288671  PMID: 6246263

Abstract

The structural polypeptides of foot-and-mouth disease virus were digested with Staphylococcus aureus V8 protease in the presence of sodium dodecyl sulfate. The protease-resistant peptides derived from temperature-sensitive mutants were compared with those of the wild type by electrofocusing in a polyacrylamide gel. Covariation between the charge shifts of different peptides indicated that they shared common sequences: only five independent peptides in all were derived from VP1, VP2, and VP3, accounting for approximately 50% of the polypeptide sequences. In two instances, amino acid substitutions that caused similar shifts in the isoelectric point were found to be located in different peptides. However, 15 mutants that possessed identical shifts in VP2 could not be distinguished by peptide analysis. The polypeptides of revertants able to grow at the nonpermissive temperature were compared with those of the parental mutants. By this test, 6 of the 12 distinguishable classes of coat protein mutations were found to covary with temperature sensitivity. In addition to true revertants, several phenotypic revertants which possessed a second charge change, either in a different structural polypeptide or in a different region of the same polypeptide, were isolated. The orientation of the recombination map was deduced from the loci of the coat protein mutations.

Full text

PDF
67

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burroughs J. N., Rowlands D. J., Sangar D. V., Talbot P., Brown F. Further evidence for multiple proteins in the foot-and-mouth disease virus particle. J Gen Virol. 1971 Oct;13(1):73–84. doi: 10.1099/0022-1317-13-1-73. [DOI] [PubMed] [Google Scholar]
  2. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  3. Cooper P. D. A genetic map of poliovirus temperature-sensitive mutants. Virology. 1968 Aug;35(4):584–596. doi: 10.1016/0042-6822(68)90287-0. [DOI] [PubMed] [Google Scholar]
  4. Doel T. R., Sangar D. V., Rowlands D. J., Brown F. A re-appraisal of the biochemical map of foot-and-mouth disease virus RNA. J Gen Virol. 1978 Nov;41(2):395–404. doi: 10.1099/0022-1317-41-2-395. [DOI] [PubMed] [Google Scholar]
  5. Harris T. J., Brown F. The location of the ploy(C) tract in the RNA of foot-and-mouth disease virus. J Gen Virol. 1976 Dec;33(3):493–501. doi: 10.1099/0022-1317-33-3-493. [DOI] [PubMed] [Google Scholar]
  6. Jarvik J., Botstein D. Conditional-lethal mutations that suppress genetic defects in morphogenesis by altering structural proteins. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2738–2742. doi: 10.1073/pnas.72.7.2738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. King A. M., Newman J. W. Temperature-sensitive mutants of foot-and-mouth disease virus with altered structural polypeptides. I. Identification by electrofocusing. J Virol. 1980 Apr;34(1):59–66. doi: 10.1128/jvi.34.1.59-66.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Lake J. R., Priston A. J., Slade W. R. A genetic recombination map of foot-and-mouth disease virus. J Gen Virol. 1975 Jun;27(3):355–367. doi: 10.1099/0022-1317-27-3-355. [DOI] [PubMed] [Google Scholar]
  10. Lake J., Mackenzie J. S. Improved technique for the isolation of temperature-sensitive mutants of foot-and-mouth disease virus. J Virol. 1973 Sep;12(3):665–668. doi: 10.1128/jvi.12.3.665-668.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MacKenzie J. S., Slade W. R., Lake J., Priston R. A., Bisby J., Laing S., Newman J. Temperature-sensitive mutants of foot-and-mouth disease virus: the isolation of mutants and observations on their properties and genetic recombination. J Gen Virol. 1975 Apr;27(1):61–70. doi: 10.1099/0022-1317-27-1-61. [DOI] [PubMed] [Google Scholar]
  12. McCahon D., Cooper P. D. Identification of poliovirus temperature-sensitive mutants having defects in virus structural protein. J Gen Virol. 1970 Jan;6(1):51–62. doi: 10.1099/0022-1317-6-1-51. [DOI] [PubMed] [Google Scholar]
  13. McCahon D., Slade W. R., Priston R. A., Lake J. R. An extended genetic recombination map for foot-and-mouth diseases virus. J Gen Virol. 1977 Jun;35(3):555–565. doi: 10.1099/0022-1317-35-3-555. [DOI] [PubMed] [Google Scholar]
  14. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  15. Sangar D. V., Black D. N., Rowlands D. J., Brown F. Biochemical mapping of the foot-and-mouth disease virus genome. J Gen Virol. 1977 May;35(2):281–297. doi: 10.1099/0022-1317-35-2-281. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES