1887

Abstract

Summary: An mRNA-dependent cell-free translation system has been developed from the human pathogenic fungus using either S30 or S100 lysates prepared from glass-bead-disrupted whole cells. Translation of the synthetic template poly(U) in this system is highly efficient at temperatures up to 37 °C and is ATP-dependent. Studies using a range of elongation-specific inhibitors suggest that the mechanism of translational elongation in is similar to that of another yeast, . A micrococcal-nuclease-treated S100 lysate was able to translate exogenously-supplied homologous mRNAs, and a range of heterologous natural mRNAs, using an initiation mechanism that is inhibited by the antibiotic edeine and the 5′ cap analogue 7-methylguanosine 5′-monophosphate (mGMP). As with cell-free lysates prepared from , the lysate is unable to initiate translation upon natural mRNAs at temperatures above 20 °C.

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/content/journal/micro/10.1099/00221287-137-4-851
1991-04-01
2021-07-23
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References

  1. Clemens M. J. 1985; Translation of eukaryotic messenger RNA in cell-free extracts. Transcription and Translation: a Practical Approach231–270 Harnes B. D., Higgins S. J. Oxford: IRL Press;
    [Google Scholar]
  2. Fried H. M., Warner J. R. 1982; Molecular cloning and analysis of the yeast gene for cycloheximide resistance and ribosomal protein L29. Nucleic Acids Research 10:3133–3148
    [Google Scholar]
  3. Gasior E., Herrera F., Sadnik I., McLaughlin C. S., Moldave K. 1979; The preparation and characterization of a cell-free system from Saccharomyces cerevisiae that translates natural messenger ribonucleic acid. Journal of Biological Chemistry 254:2965–2969
    [Google Scholar]
  4. Herrera F., Gasior E., McLaughlin C. S., Moldave K. 1979; Evidence for a nuclease in Saccharomyces cerevisiae that effects the cell-free translation of natural messenger RNA. Biochemical and Biophysical Research Communications 88:1263–1270
    [Google Scholar]
  5. Herrera F., Martinez J. A., Moreno N., Sadnik I., McLaughlin C. S., Feinberg B., Moldave K. 1984; Identification of an altered elongation factor in temperature sensitive mutant ts7′-14 of Saccharomyces cerevisiae. Journal of Biological Chemistry 259:14347–14349
    [Google Scholar]
  6. Hofbauer R., Fessl F., Hamilton B., Ruis H. 1982; Preparation of a mRNA-dependent cell-free translation system from whole cells of Saccharomyces cerevisiae. European Journal of Biochemistry 122:199–203
    [Google Scholar]
  7. Hussain I., Leibowitz M. J. 1986; Translation of homologous and heterologous messenger RNAs in a yeast cell-free system. Gene 45:13–23
    [Google Scholar]
  8. Hutchinson J. S., Feinberg B., Rothwell T. C., Moldave K. 1984; Monoclonal antibody specific for yeast elongation factor 3. Biochemistry 23:3055–3063
    [Google Scholar]
  9. Kurtz M. B., Kirsch D. R., Kelly R. 1988; The molecular genetics of Candida albicans. Microbiological Sciences 5:58–63
    [Google Scholar]
  10. Little P. F. R., Jackson I. J. 1987; Application of plasmids containing promoters specific for phage-encoded RNA polymerases. DNA Cloning: A Practical Approach 31–18 Glover D. M. Oxford: IRL Press;
    [Google Scholar]
  11. Magee P. T., Rikkerink E. H. A., Magee B. B. 1988; Methods for the genetics and molecular biology of Candida albicans. Analytical Biochemistry 175:361–372
    [Google Scholar]
  12. Mandel T., Trachsel H. 1989; Yeast, Saccharomyces cerevisiae, cell-free translation: the inhibition of high temperature is reversible. Biochimica et Biophysica Acta 1007:80–83
    [Google Scholar]
  13. Moldave K. 1985; Eukaryotic protein synthesis. Annual Review of Biochemistry 54:1109–1149
    [Google Scholar]
  14. Odds F. C. 1988 Candida and Candidosis, 2. London: Baillière Tindall;
    [Google Scholar]
  15. Pelham H. R. B., Jackson R. J. 1976; An efficient mRNA-dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67:247–256
    [Google Scholar]
  16. Rao S. S., Grollman A. P. 1967; Cycloheximide resistance in yeast: a property of the 60S ribosomal subunit. Biochemical and Biophysical Research Communications 29:696–704
    [Google Scholar]
  17. Santos M., Colthurst D. R., Wills N., McLaughlin C. S., Tuite M. F. 1990; Efficient translation of the UAG termination codon in Candida species. Current Genetics 17:487–491
    [Google Scholar]
  18. Sissons C. H. 1978; Method for yeast protein synthesis in a cell-free system. Methods in Cell Biology 20:83–99
    [Google Scholar]
  19. Skogerson L. 1979; Separation and characterization of yeast elongation factors. Methods in Enzymology 60:676–685
    [Google Scholar]
  20. Skogerson L., Wakatama E. 1976; A ribosome-dependent GTPase from yeast distinct from elongation factor 2. Proceedings of the National Academy of Sciences of the United States of America 7373–76
    [Google Scholar]
  21. Skuzeski J. M., Nichols L. M., Gesteland R. F. 1990; Analysis of leaky viral translation termination codons in vivo by transient expression of improved β-glucuronidase vectors. Plant Molecular Biology 15:65–79
    [Google Scholar]
  22. Tuite M. F., Plesset J. 1986; mRNA-dependent yeast cell-free translation systems: theory and practice. Yeast 2:35–52
    [Google Scholar]
  23. Tuite M. F., Plesset J., Moldave K., McLaughlin C. S. 1980; Faithful and efficient translation of homologous and heterologous mRNAs in an mRNA-dependent cell-free system from Saccharomyces cerevisiae. Journal of Biological Chemistry 255:8761–8766
    [Google Scholar]
  24. Uritani M., Miyazaki M. 1988; Characterization of the ATPase and GTPase activities of elongation factor 3 (EF-3). purified from yeasts. Journal of Biochemistry 103:522–530
    [Google Scholar]
  25. Vazques D., Jiminez A. 1980 Ribosomes: Structure, Function and Genetics847–869 Chambliss G., Graven G. R., Davies J. E., Davis K., Kahan L., Nomura M. Baltimore: University Park Press;
    [Google Scholar]
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