Identification of the nucleic acid binding domain of the rotavirus VP2 protein Free

Abstract

The bovine rotavirus VP2 protein is the major component of the core and forms the most internal layer surrounding the dsRNA genome. We have constructed recombinant baculoviruses expressing truncated VP2 proteins. The nucleic acid binding activity of these truncated proteins was tested by North-Western blotting experiments with single-stranded and double-stranded probes. The nucleic acid binding domain in VP2 was localized between amino acids 1 to 132. Recombinant proteins bound single-stranded and double-stranded nucleic acids, but showed less affinity for doublestranded RNA and DNA. Interactions of VP2 with the genome were investigated in viral single-shelled particles by u.v.-cross-linking. In these experiments, only VP2 protein bound the genomic RNA in purified singleshelled particles.

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1994-12-01
2024-03-28
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References

  1. Boyle J. F., Holmes K. V. 1986; RNA-binding proteins of bovine rotavirus. Journal of Virology 58:561–568
    [Google Scholar]
  2. Brémont M., Juste-Lesage P., Chabanne-Vautherot D., Char-Pilienne A., Cohen J. 1992; Sequences of the four larger proteins of a porcine group C rotavirus and comparison with the equivalent group A rotavirus proteins. Virology 168:684–692
    [Google Scholar]
  3. Brüssow H., Bruttin A., Marc-Martin S. 1990; Polypeptide composition of the rotavirus empty capsids and their possible use as a subunit vaccine. Journal of Virology 64:3635–3642
    [Google Scholar]
  4. Cohen J. 1977; Ribonucleic acid polymerase associated with purified calf rotavirus. Journal of General Virology 36:395–402
    [Google Scholar]
  5. Ernst H., Duhl J. A. 1989; Nucleotide sequence of genomic segment 2 of human rotavirus Wa. Nucleic Acids Research 17:4382
    [Google Scholar]
  6. Estes M. K., Cohen J. 1989; Rotavirus gene structure and function. Microbiological Reviews 53:410–449
    [Google Scholar]
  7. Fox J. M., Johnson J. E., Young M. J. 1994; RNA/protein interactions in icosahedral virus assembly. Seminars in Virology 5:51–60
    [Google Scholar]
  8. Garnier J., Osguthorpe D. J., Robson B. 1978; Analysis of the accuracy and implications of the simple methods for predicting secondary structure of globular proteins. Journal of Molecular Biology 120:97–120
    [Google Scholar]
  9. Kapahnke R., Rappold W., Desselberger U., Riesner D. 1986; The stiffness of dsRNA: hydrodynamic studies on fluorescence- labelled RNA segments of bovine rotavirus. Nucleic Acids Research 14:3215–3228
    [Google Scholar]
  10. Kitts P. A., Ayres M. D., Possee R. D. 1990; Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Research 18:5667–5672
    [Google Scholar]
  11. Kumar A., Charpilienne A., Cohen J. 1989; Nucleotide sequence of the gene encoding for the RNA binding protein (VP2) of RF bovine rotavirus. Nucleic Acids Research 17:21–26
    [Google Scholar]
  12. Kunkel T. A. 1985; Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences U.S.A.: 82488
    [Google Scholar]
  13. Labbé M., Charpilienne A., Crawford S. E., Estes M. K., Cohen J. 1991; Expression of rotavirus VP2 produces empty corelike particles. Journal of Virology 65:2946–2952
    [Google Scholar]
  14. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  15. L’Haridon R., Scherrer R. 1976; In vitro culture of a rotavirus associated with neonatal calf scours. Annales de Recherche Veteri- naires 7:373–381
    [Google Scholar]
  16. Lindsay A. D., Vonderfecht S. L., Eiden J. J. 1994; Group B rotavirus VP2: sequence analysis, expression and gene coding assignment. Virology 199:141–150
    [Google Scholar]
  17. Liu M., Mattion M. N., Estes M. K. 1992; Rotavirus VP3 expressed in insect cells possesses guanylyltransferase activity. Virology 188:77–84
    [Google Scholar]
  18. Loudon P. T., Roy P. 1992; Interaction of nucleic acids with corelike and subcore-like particles of bluetongue virus. Virology 191:231–236
    [Google Scholar]
  19. McCormack S. J., Thomas D. C., Samuel C. E. 1992; Mechanism of interferon action: identification of a RNA binding domain within the N-terminal region of the human RNA-dependent PI/elF- 2a protein kinase. Virology 188:47–56
    [Google Scholar]
  20. Mitchell D. B., Both G. W. 1990; Completion of the genomic sequence of the simian rotavirus SA11: nucleotide sequences of segments 1, 2 and 3. Virology 177:324–331
    [Google Scholar]
  21. Pizarro J. L., Sandino A. M., Pizarro J. M., Fernández J., Spencer E. 1991; Characterization of rotavirus guanylyltransferase activity associated with polypeptide VP3. Journal of General Virology 72:325–332
    [Google Scholar]
  22. Prasad B. V. V., Wang G. J., Clerx J. P. M., Chiu W. 1988; Three-dimensional structure of rotavirus. Journal of Molecular Biology 199:269–275
    [Google Scholar]
  23. Roseto A., Scherrer R., Cohen J., Guillemin M. C., Charpilienne A., Feynerol C., Peries J. 1983; Isolation and characterization of anti-rotavirus immunoblobulins secreted by cloned hybridoma cells lines. Journal of General Virology 64:237–240
    [Google Scholar]
  24. Schägger H., Von Jagow G. 1987; Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analytical Biochemistry 166:368–379
    [Google Scholar]
  25. Stewart L., Schatz G., Vogt V. M. 1990; Properties of avian retrovirus particles defective in viral protease. Journal of Virology 64:5076–5092
    [Google Scholar]
  26. Summers M. D., Smith G. E. 1987; A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin 1555:
    [Google Scholar]
  27. Tian Y., Tarlow O., Mccrae M. A. 1990; Nucleotide sequence of gene 2 of the UK tissue culture adapted strain of bovine rotavirus. Nucleic Acids Research 18:4015
    [Google Scholar]
  28. Valenzuela S., Pizarro J., Sandino A. M., Vasquez M., Fernandez ]., Hernandez O., Patton J., Spencer E. 1991; Photoaffinity labeling of rotavirus VP1 with 8-azido-ATP: identification of the viral RNA polymerase. Journal of Virology 65:3964–3967
    [Google Scholar]
  29. Zeng C. Q.-Y., Labbé M., Cohen C., Prasad B. V. V., Chen D., Ramig R. F., Estes M. K. 1994; Characterization of rotavirus VP2 particles. Virology 201:55–65
    [Google Scholar]
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