1887

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Volume 70, Issue 9

Isolation and Characterization of Canine Distemper Virus Nucleocapsid Variants Free

Abstract

Summary

Nucleocapsid (NC) variants expressed by the Onderstepoort strain of canine distemper virus (CDV) were ultrastructurally and biochemically characterized. Three isolated variants were defined which corresponded to the three variants observed within the cytoplasm of infected cells. Dense NC (D-NC), isolated on discontinuous caesium chloride (CsCl) isopycnic gradients, had an average density of 1·2971 ± 0·0042 g/ml. Ultrastructurally, D-NC were 1620·0 ± 112·1 nm in length with a 20·1 ± 1·3 nm outer and a 5·8 ± 0·7 nm inner core diameter. The D-NC protein composition was 89·7% of a 61K protein (N), 8·4% of a 75K protein (P) and 1·9% of a 160K to 200K protein (L). A single species of nucleic acid, 15 kb in length, was isolated from D-NC. Light NC (L-NC), similarly isolated, had an average density of 1·2894 ± 0·0040 g/ml. L-NC differed ultrastructurally from D-NC in that poor resolution of NC subunits, a larger outer diameter (32·0 ± 2·8 nm), and a greater inner core diameter (10·4 ± 0·6 nm) were observed. The average L-NC strand length was 1574·4 ± 115·8 nm. The protein composition was the same as D-NC with the exception of an additional 70K protein, representing 4·0 to 7·7% of the total L-NC protein mass. A 15 kb nucleic acid was also identified in L-NC, although heightened sensitivity of encapsidated L-NC nucleic acid to non-specific nuclease degradation was observed. The ratio of D-NC to L-NC isolated from individual virus preparations varied and was independent of viral infectivity. A third NC variant, defective-NC (Df-NC), was also identified. This had the lowest density on CsCl gradients (1·2460 ± 0·0046 g/ml). The Df-NC structures were truncated to a uniform length of 87·0 ± 5·8 nm. Diameter measurements were between those of D-NC and L-NC, being 24·4 ± 1·4 nm (outer) and 6·9 ± 0·4 nm (inner core). Like L-NC, the 70K protein was present but in greater amounts, representing as much as 43·7% of the total Df-NC protein mass. RNase A-sensitive nucleic acid was isolated from Df-NC which ranged in size from 1·16 to 0·67 kb with a majority of the material being 0·86 kb in length. For both L-NC and Df-NC, canine CDV convalescent serum reacted with viral N and P proteins in Western blot analyses but not with the 70K protein, suggesting a host cellular origin for the latter.

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Keyword(s): CDV and nucleocapsid
© Society for General Microbiology 1989
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1989-09-01
2024-04-25
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References

  1. Bohn W., Rutter G., Hohenberg H., Mannweiler K., Nobis P. 1986; Involvement of actin filaments in budding of measles virus: studies on cytoskeletons of infected cells.. Virology 149:91–106
     [Google Scholar]
  2. Bruner R., Vinograd J. 1965; The evaluation of standard sedimentation coefficients of sodium RNA and sodium DNA from sedimentation velocity data in concentrated NaCl and CsCl solutions.. Biochimica et biophysica acta 108:18–29
     [Google Scholar]
  3. Bussell R. H., Waters D. J., Seals M. K. 1974; Measles, canine distemper and respiratory syncytial virions and nucleocapsids. A comparative study of their structure, polypeptide and nucleic acid composition.. Medical Microbiology and Immunology 160:105–124
     [Google Scholar]
  4. Cattaneo R., Rebmann G., Schmidt A., Baczko K., , TER Meulen V., Billeter M. A. 1987; Altered transcription of a defective measles virus genome derived from a diseased human brain.. EMBO Journal 6:681–688
     [Google Scholar]
  5. Chinchar V. G., Portner A. 1981; Inhibition of RNA synthesis following proteolytic cleavage of Newcastle disease virus P protein.. Virology 115,:192–202
     [Google Scholar]
  6. Chui L.W-L., vainionpÄÄ R., Marusyk R., Salmi A., Norrby E. 1986; Nuclear accumulation of measles virus nucleoprotein associated with a temperature-sensitive mutant.. Journal of General Virology 67:2153–2161
     [Google Scholar]
  7. Croce C. M., Linnenbach A., Hall W., Steplewski Z., Koprowski H. 1980; Production of human hybridomas secreting antibodies to measles virus.. Nature London: 288:488–489
     [Google Scholar]
  8. Deshpande K. L., Portner A. 1985; Monoclonal antibodies to the P protein of Sendai virus define its structure and role in transcription.. Virology 140:125–134
     [Google Scholar]
  9. Dubois-Dalcq M., Barbosa L. H., Hamilton R., Sever J. L. 1974; Comparison between productive and latent subacute sclerosing panencephalitis viral infection in vitro: an electron microscopic and immunoperoxidase study.. Laboratory Investigation 30,:241–250
     [Google Scholar]
  10. Finch J. T., Gibbs A. J. 1970; Observations on the structure of the nucleocapsids of some paramyxoviruses.. Journal of General Virology 6:141–150
     [Google Scholar]
  11. Fournier J., Priuvat A., Bouteille M. 1981; Structural changes in the cell nucleus during measles virus infection of cerebellar explants.. Journal of ultrastructure Research 77:319–328
     [Google Scholar]
  12. Giuffre R. M., Tovell D. R., Kay C. M., Tyrrell D. L. J. 1982; Evidence for the interaction between the membrane protein of a paramyxovirus and actin.. Journal of Virology 42,:963–968
     [Google Scholar]
  13. Kingsbury D. W. 1972; Paramyxovirus replication.. Current Topics in Microbiology and Immunology 59,:1–33
     [Google Scholar]
  14. Kingsbury D. W., Darlington R. W. 1968; Isolation and properties of Newcastle disease virus nucleocapsid.. Journal of Virology 2:248–255
     [Google Scholar]
  15. Koestner A., Long J. 1970; Ultrastructure of canine distemper virus in explant tissue cultures of canine cerebellum.. Laboratory Investigation 23:196–201
     [Google Scholar]
  16. Kolakofsky D., Blumberg B. M. 1982; A model for the control of non-segmented negative strand virus genome replication.. In Virus Persistence Mahy B. W. J., Minson A. C., Darby G. K. Cambridge:: Cambridge University Press.;
     [Google Scholar]
  17. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature London: 227,:680–685
     [Google Scholar]
  18. Lamb R. A., Mahy B. W. J., Choppin P. W. 1976; The synthesis of Sendai virus polypeptides in infected cells.. Virology 69,:116–131
     [Google Scholar]
  19. Lazzarini R. A., Keene J. D., Schubert M. 1981; The origins of defective interfering particles of the negative strand RNA viruses.. Cell 26,:145–154
     [Google Scholar]
  20. Leppert M., Kort L., Kolakofsky D. 1977; Further characterization of Sendai virus DI-RNAs: a model for their generation.. Cell 12,:539–552
     [Google Scholar]
  21. Matsumoto N. 1966; Studies on measles virus in tissue culture. II. Electron microscopic study of measles virus infected cells and localization of virus antigen examined by ferritin-conjugated antibody method.. Bulletin of the Yamaguchi Medical School 13,:167–189
     [Google Scholar]
  22. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. 1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins.. Science 211,:1437
     [Google Scholar]
  23. Miller k. 1987; Gel electrophoresis of RNA.. Focus 9:(3)14. Gaithersburg:: Bethesda Research Laboratories.;
     [Google Scholar]
  24. Minagawa T., Sakuma T., Kuwajima s., Yamamoto T. K., Iida H. 1976; Characterization of measles viruses in establishment of persistent infections in human lymphoid cell line.. Journal of General Virology 33,:361–379
     [Google Scholar]
  25. Narang H. K. 1981; Comparative morphology of measles virus and paramyxovirus-like tubules in multiple sclerosis using ruthenium red stain.. Neuropathology and Applied Neurobiology 7,:411–420
     [Google Scholar]
  26. Oglesbee M., Tatalick L., Ringler S., Rice J., Krakowka S. 1989; Rapid isolation of morbillivirus nucleocapsid for genomic RNA cDNA cloning and the production of specific core protein antisera.. Journal of Virological Methods in press
    [Google Scholar]
  27. Oyanagi S., Ter Meulen V., Muller D. 1970; Electron microscope observations in subacute sclerosing panencephalitis brain cell cultures: their correlation with cytochemical and immunocytological findings.. Journal of Virology 6,:370–379
     [Google Scholar]
  28. Oyanagi S., Ter Meulen V., Katz M., Koprowski H. 1971; Comparison of subacute sclerosing panencephalitis and measles virus: an electron microscope study.. Journal of Virology 7,:176–182
     [Google Scholar]
  29. Portner A., Kingsbury D. W. 1976; Regulatory events in the synthesis of Sendai virus polypeptides and their assembly into virions.. Virology 73,:79–88
     [Google Scholar]
  30. Portner A., Murti K. G. 1986; Localization of P, NP, and M proteins on Sendai virus nucleocapsid using immunogold labeling.. Virology 150,:469–478
     [Google Scholar]
  31. Pringle C. R. 1987; Rhabdovirus genetics.. In The Rhabdoviruses167–243 Wagner R. R. New York:: Plenum Press.;
     [Google Scholar]
  32. Re G. G., Gupta K. C., Kingsbury D. W. 1983; Genomic and copy-back 3′ termini in Sendai virus defective interfering RNA species.. Journal of Virology 45,:659–664
     [Google Scholar]
  33. Rima B. K. 1983; The proteins of morbilliviruses.. Journal of General Virology 64,:1205–1219
     [Google Scholar]
  34. Robbins S. J. 1983; Progressive invasion of cell nuclei by measles virus in persistently infected human cells.. Journal of General Virology 64,:2335–2338
     [Google Scholar]
  35. Robbins S. J., Bussell R. H. 1979; Structural phosphoproteins associated with purified measles virions and cytoplasmic nucleocapsids.. Intervirology 12,:96–102
     [Google Scholar]
  36. Robbins S. J., Bussell R. H., Rapp F. 1980; Isolation and partial characterization of two forms of cytoplasmic nucleocapsid from measles virus-infected cells.. Journal of General Virology 47,:301–310
     [Google Scholar]
  37. Rozenblatt S., Koch T., Pinhasi O., Bratosin S. 1979; Infective substructures of measles virus isolated from acutely and persistently infected cells.. Journal of Virology 32,:329–333
     [Google Scholar]
  38. Schubert M., Harmison G. G., Meier E. 1984; Primary structure of the vesicular stomatitis virus polymerase (L) gene: evidence for a high frequency of mutations.. Journal of Virology 51,:505–514
     [Google Scholar]
  39. Seifried A. S., Albrecht P., Milstein J. B. 1978; Characterization of an RNA-dependent RNA polymerase activity associated with measles virus.. Journal of Virology 25,:781–787
     [Google Scholar]
  40. Tyrrell D. L. J., Norrby E. 1978; Structural polypeptides of measles virus.. Journal of General Virology 39,:219–229
     [Google Scholar]
  41. Uchiyama T., Takeda K., Yamanouchi K. 1985; Replication of measles virus and localization of the viral antigens in long term infection of organotypic cultures of hamster dorsal root ganglion.. Japanese Journal of Experimental Medicine 55,:109–121
     [Google Scholar]
  42. Udem S. A., Cook K. A. 1984; Isolation and characterization of measles virus intracellular nucleocapsid RNA.. Journal of Virology 49,:57–65
     [Google Scholar]
  43. Waters D. J., Bussell R. H. 1974; Isolation and comparative study of the nucleocapsids of measles and canine distemper viruses from infected cells.. Virology 61,:64–79
     [Google Scholar]
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Isolation and Characterization of Canine Distemper Virus Nucleocapsid Variants
J. Gen. Virol. 70, 2409 (1989); https://doi.org/10.1099/0022-1317-70-9-2409
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