Transcription and Translation of Bluetongue Virus mRNA Free

Abstract

Summary

Fractionation of transcribed bluetongue virus (BTV) mRNA by agarose gel electrophoresis resulted in the separation of eight of the 10 species. The relative molar ratio of the mRNAs confirmed that mRNA 5 was transcribed more frequently than would be predicted from the size of the S5 genome segment, while mRNA 10 was transcribed less frequently. translation of unfractionated BTV mRNAs resulted in the synthesis of the seven known structural proteins (P1 to P7) and two known non-structural proteins (NS1 and NS2). Two additional non-structural proteins (NS3 and NS3A) with of 28K and 25K respectively were identified. The protein coding assignments for the medium- and small-sized double-stranded RNA genome segments of BTV serotype 10 were found to correspond to those reported for BTV-1 and BTV-17. The peptide maps of NS1, NS2, NS3 and NS3A synthesized corresponded to those of their counterparts synthesized in infected cells. Protein NS3A appeared to be a truncated form of NS3, since its peptide map completely overlapped that of NS3. Proteins NS3 and NS3A were present in very small amounts in the soluble fraction of the cytoplasm of infected cells, and were synthesized in variable amounts , whereas the other nine viral proteins were synthesized in constant molar ratios. A difference in the relative molar ratios in which some of the BTV proteins were synthesized and was observed. , protein NS1 was translated in the largest amount but , NS2 was the most efficiently translated protein. Conversely, protein P6 was translated much more efficiently than .

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1988-03-01
2024-03-28
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References

  1. BONNER W. M., LASKEY R. A. 1974; A film detection method for tritium-labeled proteins and nucleic acids in Polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  2. BURGE B. W., HUANG A. S. 1979; Conserved polypeptides in the proteins of vesicular stomatitis virus. Virology 95:445–452
    [Google Scholar]
  3. CLEVELAND D. W., FISCHER S. G., KIRSCHNER M. W., LAEMMLI U. K. 1977; Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. Journal of Biological Chemistry 252:1102–1106
    [Google Scholar]
  4. GRUBMAN M. J., APPLETON J. A., LETCHWORTH G. J. III 1983; Identification of bluetongue virus type 17 genome segments coding for polypeptides associated with virus neutralization and intergroup reactivity. Virology 131:355–366
    [Google Scholar]
  5. HUISMANS H. 1979; Protein synthesis in bluetongue infected cells. Virology 92:385–396
    [Google Scholar]
  6. HUISMANS H., VERWOERD D. W. 1973; Control of transcription during expression of the bluetongue virus genome. Virology 52:81–88
    [Google Scholar]
  7. HUISMANS H, BREMER C. W., BARBER T. L. 1979; The nucleic acid and proteins of epizootic haemorrhagic disease virus. Onderstepoort Journal of Veterinary Research 46:95–104
    [Google Scholar]
  8. HUISMANS H., VAN DER WALT N. T., CLOETE M., ERASMUS B. J. 1987a; Isolation of a capsid protein of bluetongue virus that induces a protective immune response in sheep. Virology 157:172–179
    [Google Scholar]
  9. HUISMANS H., VAN DIJK A. A., BAUSKIN A. R. 1987b; In vitro phosphorylation and purification of a nonstructural protein of bluetongue virus with affinity for single-stranded RNA. Journal of Virology 61:3589–3595
    [Google Scholar]
  10. LAEMMLI U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  11. LEE J. W., ROY P. 1986; Nucleotide sequence of acDNA clone of RNA segment 10 of bluetongue virus (serotype 10). Journal of General Virology 67:2833–2837
    [Google Scholar]
  12. LEVIN K. H., SAMUEL C. E. 1980; Biosynthesis of reovirus-specified polypeptides. Virology 106:1–13
    [Google Scholar]
  13. LOENING U. E. 1967; The fractionation of high molecular weight ribonucleic acid by Polyacrylamide gel electrophoresis. Biochemical Journal 102:251–257
    [Google Scholar]
  14. McCrae M. A., JOKLIK W. K. 1978; The nature of the polypeptide encoded by each of the double-stranded RNA segments of reovirus type 3. Virology 89:578–593
    [Google Scholar]
  15. MERTENS P. P. C, BROWN F., SANGAR D. V. 1984; Assignment of the genome segments of bluetongue virus type 1 to the proteins which they encode. Virology 135:207–217
    [Google Scholar]
  16. MERTENS P. P. C, BURROUGHS J. N., ANDERSON J. 1987; Purification and properties of virus particles, infectious subviral particles, and cores of bluetongue virus serotypes 1 and 4. Virology 157:375–386
    [Google Scholar]
  17. NAMIKI M., SUZUKI Y., NAKAGAWA S., YOSHIOKA I., SAITO Y. 1983; Characterization of ribonucleic acid transcriptase in Ibaraki virus core particles. Journal of General Virology 64:1397–1400
    [Google Scholar]
  18. NEELEMAN L., VAN VLOTEN-DOTING L. 1978; Excess of micrococcal nuclease may harm the exogenous mRNA dependent rabbit reticulocyte cell-free system. FEBS Letters 95:103–106
    [Google Scholar]
  19. 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]
  20. RAE B. P., ELLIOTT R. M. 1986; Characterization of the mutations responsible for the electrophoretic mobility differences in the NS proteins of vesicular stomatitis virus New Jersey complementation group E mutants. Journal of General Virology 67:2635–2643
    [Google Scholar]
  21. RANU R. S., LONDON I. M. 1979; Regulation of protein synthesis in rabbit reticulocyte lysates; preparation of efficient protein synthesis lysates and the purification and characterization of the heme regulated translational inhibitory protein kinase. Methods in Enzymology 60:459–484
    [Google Scholar]
  22. VAN DIJK A. A., HUISMANS H. 1980; The in vitro activation and further characterization of the bluetongue virus-associated transcriptase. Virology 104:347–356
    [Google Scholar]
  23. VERWOERD D. W., OELLERMANN R. A., BROEKMAN J., WEISS K. 1967; The serological relationship of South African bovine enterovirus strains (Ebco SA-I and II) and the growth characteristics in cell culture of the prototype strain (Ebco SA-I). Onderstepoort Journal of Veterinary Research 34:41–52
    [Google Scholar]
  24. VERWOERD D. W., ELS H. J., DE VILIERS E.-M., HUISMANS H. 1972; Structure of the bluetongue virus capsid. Journal of Virology 10:783–794
    [Google Scholar]
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