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. 1981 Jan;37(1):256–267. doi: 10.1128/jvi.37.1.256-267.1981

Identification of the P proteins and other disulfide-linked and phosphorylated proteins of Newcastle disease virus.

G W Smith, L E Hightower
PMCID: PMC171003  PMID: 7218425

Abstract

A unique abundant protein, designated P by analogy to the putative polymerase proteins of other paramyxoviruses, was identified in purified Newcastle disease virus. Under nonreducing conditions the P proteins could be separated from other viral proteins on sodium dodecyl sulfate-polyacrylamide gels. The P proteins were isolated from detergent-solubilized virions as 53,000- to 55,000-dalton monomers and disulfide-linked trimers. Distinct forms of P having four different isoelectric points and two different electrophoretic mobilities were resolved by two-dimensional electrophoresis. Two forms of P were phosphorylated, as were the nucleocapsid protein and non-glycosylated membrane protein. In addition to disulfide-linked forms of P, dimers of the hemagglutinin-neuraminidase glycoprotein and two disulfide-linked versions of the fusion glycoprotein were identified. Several electrophoretic variants of the nucleocapsid protein that were probably created by intrachain disulfide bonding were also isolated from virions under nonreducing conditions. The locations of the newly identified proteins were determined by detergent-salt fractionation of virions and by surface-selective radioiodination of the viral envelope. The P proteins were associated with nucleocapsids and were not detected at the surface of virions. Both forms of the fusion glycoproteins were on the exterior of the viral envelope. Herein the properties of the P proteins are compared with similar proteins of rhabdoviruses and other paramyxoviruses, and a role for multiple forms of proteins in the genetic economy of newcastle disease virus is discussed.

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Selected References

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  1. Abraham G., Banerjee A. K. Sequential transcription of the genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 May;73(5):1504–1508. doi: 10.1073/pnas.73.5.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ball L. A., White C. N. Order of transcription of genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):442–446. doi: 10.1073/pnas.73.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bell J. C., Prevec L. Proteins of vesicular stomatitis virus. V. Identification of a precursor to the phosphoprotein of Piry virus. J Virol. 1979 Apr;30(1):56–63. doi: 10.1128/jvi.30.1.56-63.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buetti E., Choppin P. W. The transcriptase complex of the paramyxovirus SV5. Virology. 1977 Oct 15;82(2):493–508. doi: 10.1016/0042-6822(77)90021-6. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Clinkscales C. W., Bratt M. A., Morrison T. G. Synthesis of Newcastle disease virus polypeptides in a wheat germ cell-free system. J Virol. 1977 Apr;22(1):97–101. doi: 10.1128/jvi.22.1.97-101.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clinton G. M., Burge B. W., Huang A. S. Effects of phosphorylation and pH on the association of NS protein with vesicular stomatitis virus cores. J Virol. 1978 Aug;27(2):340–346. doi: 10.1128/jvi.27.2.340-346.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collins P. L., Hightower L. E., Ball L. A. Transcription and translation of Newcastle disease virus mRNA's in vitro. J Virol. 1978 Oct;28(1):324–336. doi: 10.1128/jvi.28.1.324-336.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Collins P. L., Hightower L. E., Ball L. A. Transcriptional map for Newcastle disease virus. J Virol. 1980 Sep;35(3):682–693. doi: 10.1128/jvi.35.3.682-693.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Colonno R. J., Stone H. O. Isolation of a transcriptive complex from Newcastle disease virions. J Virol. 1976 Sep;19(3):1080–1089. doi: 10.1128/jvi.19.3.1080-1089.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Colonno R. J., Stone H. O. Methylation of messenger RNA of Newcastle disease virus in vitro by a virion-associated enzyme. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2611–2615. doi: 10.1073/pnas.72.7.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Colonno R. J., Stone H. O. Newcastle disease virus mRNA lacks 2'-O-methylated nucleotides. Nature. 1976 Jun 17;261(5561):611–614. doi: 10.1038/261611a0. [DOI] [PubMed] [Google Scholar]
  13. Goldman E., Lodish H. F. Inhibition of replication of ribonucleic acid bacteriophage f2 by superinfection with bacteriophage T4. J Virol. 1971 Oct;8(4):417–429. doi: 10.1128/jvi.8.4.417-429.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Henning R., Milner R. J., Reske K., Cunningham B. A., Edelman G. M. Subunit structure, cell surface orientation, and partial amino-acid sequences of murine histocompatibility antigens. Proc Natl Acad Sci U S A. 1976 Jan;73(1):118–122. doi: 10.1073/pnas.73.1.118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hightower L. E., Bratt M. A. Protein synthesis in Newcastle disease virus-infected chicken embryo cells. J Virol. 1974 Apr;13(4):788–800. doi: 10.1128/jvi.13.4.788-800.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hightower L. E., Morrison T. G., Bratt M. A. Relationships among the polypeptides of Newcastle disease virus. J Virol. 1975 Dec;16(6):1599–1607. doi: 10.1128/jvi.16.6.1599-1607.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huang A. S., Baltimore D., Bratt M. A. Ribonucleic acid polymerase in virions of Newcastle disease virus: comparison with the vesicular stomatitis virus polymerase. J Virol. 1971 Mar;7(3):389–394. doi: 10.1128/jvi.7.3.389-394.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kingsbury D. W., Darlington R. W. Isolation and properties of Newcastle disease virus nucleocapsid. J Virol. 1968 Mar;2(3):248–255. doi: 10.1128/jvi.2.3.248-255.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kingsbury D. W., Hsu C. H., Murti K. G. Intracellular metabolism of sendai virus nucleocapside. Virology. 1978 Nov;91(1):86–94. doi: 10.1016/0042-6822(78)90357-4. [DOI] [PubMed] [Google Scholar]
  20. Kingsford L., Emerson S. U. Transcriptional activities of different phosphorylated species of NS protein purified from vesicular stomatitis virions and cytoplasm of infected cells. J Virol. 1980 Mar;33(3):1097–1105. doi: 10.1128/jvi.33.3.1097-1105.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Lamb R. A., Choppin P. W. The synthesis of Sendai virus polypeptides in infected cells. III. Phosphorylation of polypeptides. Virology. 1977 Sep;81(2):382–397. doi: 10.1016/0042-6822(77)90154-4. [DOI] [PubMed] [Google Scholar]
  23. Lamb R. A. The phosphorylation of sendai virus proteins by a virus particle-associated protein kinase. J Gen Virol. 1975 Mar;26(3):249–263. doi: 10.1099/0022-1317-26-3-249. [DOI] [PubMed] [Google Scholar]
  24. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  25. Li J. K., Miyakawa T., Fox C. F. Protein organization in Newcastle disease virus as revealed by perturbant treatment. J Virol. 1980 Apr;34(1):268–271. doi: 10.1128/jvi.34.1.268-271.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Markwell M. A., Fox C. F. Protein-protein interactions within paramyxoviruses identified by native disulfide bonding or reversible chemical cross-linking. J Virol. 1980 Jan;33(1):152–166. doi: 10.1128/jvi.33.1.152-166.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  28. Marx P. A., Portner A., Kingsbury D. W. Sendai virion transcriptase complex: polyeptide composition and inhibition by virion envelope proteins. J Virol. 1974 Jan;13(1):107–112. doi: 10.1128/jvi.13.1.107-112.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Meager A., Burke D. C. Studies on the structural basis of the RNA polymerase activity of newcastle disease virus particles. J Gen Virol. 1973 Mar;18(3):305–317. doi: 10.1099/0022-1317-18-3-305. [DOI] [PubMed] [Google Scholar]
  30. Moore N. F., Burke D. C. Characterization of the structural proteins of different strains of Newcastle disease virus. J Gen Virol. 1974 Nov;25(2):275–289. doi: 10.1099/0022-1317-25-2-275. [DOI] [PubMed] [Google Scholar]
  31. Mountcastle W. E., Compans R. W., Caliguiri L. A., Choppin P. W. Nucleocapsid protein subunits of simian virus 5, Newcastle disease virus, and Sendai virus. J Virol. 1970 Nov;6(5):677–684. doi: 10.1128/jvi.6.5.677-684.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mountcastle W. E., Compans R. W., Choppin P. W. Proteins and glycoproteins of paramyxoviruses: a comparison of simian virus 5, Newcastle disease virus, and Sendai virus. J Virol. 1971 Jan;7(1):47–52. doi: 10.1128/jvi.7.1.47-52.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nagai Y., Klenk H. D. Activation of precursors to both glycoporteins of Newcastle disease virus by proteolytic cleavage. Virology. 1977 Mar;77(1):125–134. doi: 10.1016/0042-6822(77)90412-3. [DOI] [PubMed] [Google Scholar]
  34. Nagai Y., Ogura H., Klenk H. Studies on the assembly of the envelope of Newcastle disease virus. Virology. 1976 Feb;69(2):523–538. doi: 10.1016/0042-6822(76)90482-7. [DOI] [PubMed] [Google Scholar]
  35. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  36. Ozawa M., Asano A., Okada Y. Importance of interpeptide disulfide bond in a viral glycoprotein with hemagglutination and neuraminidase activities. FEBS Lett. 1976 Nov;70(1):145–149. doi: 10.1016/0014-5793(76)80745-4. [DOI] [PubMed] [Google Scholar]
  37. Raghow R., Kingsbury D. W., Portner A., George S. Topography of a flexible ribonucleoprotein helix: protein-protein contacts in Sendai virus nucleocapsids. J Virol. 1979 Jun;30(3):701–710. doi: 10.1128/jvi.30.3.701-710.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Raghow R., Portner A., Hsu C. H., Clark S. B., Kingsbury D. W. Charge heterogeneity in polypeptides of negative strand RNA viruses. Virology. 1978 Oct 15;90(2):214–225. doi: 10.1016/0042-6822(78)90305-7. [DOI] [PubMed] [Google Scholar]
  39. Samson A. C., Chambers P., Dickinson J. H. Location of post-translational cleavage events within F and HN glycoproteins of Newcastle disease virus. J Gen Virol. 1980 Mar;47(1):19–27. doi: 10.1099/0022-1317-47-1-19. [DOI] [PubMed] [Google Scholar]
  40. Scheid A., Choppin P. W. Isolation and purification of the envelope proteins of Newcastle disease virus. J Virol. 1973 Feb;11(2):263–271. doi: 10.1128/jvi.11.2.263-271.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Scheid A., Choppin P. W. Two disulfide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology. 1977 Jul 1;80(1):54–66. doi: 10.1016/0042-6822(77)90380-4. [DOI] [PubMed] [Google Scholar]
  42. Seto J. T., Becht H., Rott R. Effect of specific antibodies on biological functions of the envelope components of Newcastle disease virus. Virology. 1974 Oct;61(2):354–360. doi: 10.1016/0042-6822(74)90273-6. [DOI] [PubMed] [Google Scholar]
  43. Smith G. W., Schwalbe J. C., Hightower L. E. Maturation of the hemagglutinin-neuraminidase and fusion glycoproteins of two biologically distinct strains of Newcastle disease virus. Prog Clin Biol Res. 1980;40:275–291. [PubMed] [Google Scholar]
  44. Sokol F., Clark H. F. Phosphoproteins, structural components of rhabdoviruses. Virology. 1973 Mar;52(1):246–263. doi: 10.1016/0042-6822(73)90413-3. [DOI] [PubMed] [Google Scholar]
  45. Stone H. O., Kingsbury D. W., Darlington R. W. Sendai virus-induced transcriptase from infected cells: polypeptides in the transcriptive complex. J Virol. 1972 Nov;10(5):1037–1043. doi: 10.1128/jvi.10.5.1037-1043.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  47. Weiss S. R., Bratt M. A. Polyadenylate sequences on Newcastle disease virus mRNA synthesized in vivo and in vitro. J Virol. 1974 Jun;13(6):1220–1230. doi: 10.1128/jvi.13.6.1220-1230.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zaides V. M., Selimova L. M., Zhirnov O. P., Bukrinskaya A. G. Protein synthesis in Sendai virus-infected cells. J Gen Virol. 1975 Jun;27(3):319–327. doi: 10.1099/0022-1317-27-3-319. [DOI] [PubMed] [Google Scholar]

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