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. 1981 Jun;38(3):840–848. doi: 10.1128/jvi.38.3.840-848.1981

Characterization of polypeptides immunoprecipitable from Pichinde virus-infected BHK-21 cells.

D G Harnish, W C Leung, W E Rawls
PMCID: PMC171221  PMID: 6264158

Abstract

Using hamster anti-Pichinde virus serum, we immunoprecipitated polypeptides from BHK-21 cells infected with Pichinde virus. Seven immunoprecipitable polypeptides exhibited a time- and multiplicity of infection-dependent appearance when the cultures were pulse-labeled with L-[35S]methionine for 1 h. The predominant polypeptide was a nucleoprotein (NP) of 64,000 daltons. Components of 48,000, 38,000, and 28,000 daltons, when analyzed by two-dimensional tryptic peptide mapping, were found to be derived from NP. After a 3-h chase period, polypeptides of 17,000, 16,500, and 14,000 daltons were evident, and peptide mapping revealed that these three polypeptides were also related to NP. During a series of pulse-chase experiments, a 79,000-dalton glycoprotein (GPC) was cleaved to glycoproteins of 52,000 and 36,000 daltons. Radiolabel in a polypeptide of approximately 200,000 daltons (L) did not chase into smaller cleavage products. L, GPC, and NP were found to be unique by two-dimensional tryptic peptide mapping. Comparison of polypeptides immunoprecipitated from infected cells with structural components of purified virus revealed that L protein was evident in both. This is the first report of a high-molecular-weight polypeptide in Pichinde virus particles and infected cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Buchmeier M. J., Gee S. R., Rawls W. E. Antigens of Pichinde virus I. Relationship of soluble antigens derived from infected BHK-21 cells to the structural components of the virion. J Virol. 1977 Apr;22(1):175–186. doi: 10.1128/jvi.22.1.175-186.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchmeier M. J., Oldstone M. B. Protein structure of lymphocytic choriomeningitis virus: evidence for a cell-associated precursor of the virion glycopeptides. Virology. 1979 Nov;99(1):111–120. doi: 10.1016/0042-6822(79)90042-4. [DOI] [PubMed] [Google Scholar]
  4. Carter M. F., Biswal N., Rawls W. E. Characterization of nucleic acid of pichinde virus. J Virol. 1973 Jan;11(1):61–68. doi: 10.1128/jvi.11.1.61-68.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dobos P., Rowe D. Peptide map comparison of infectious pancreatic necrosis virus-specific polypeptides. J Virol. 1977 Dec;24(3):805–820. doi: 10.1128/jvi.24.3.805-820.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Lennarz W. J. Lipid linked sugars in glycoprotein synthesis. Science. 1975 Jun 6;188(4192):986–991. doi: 10.1126/science.167438. [DOI] [PubMed] [Google Scholar]
  8. Leung W. C., Ghosh H. P., Rawls W. E. Strandedness of Pichinde virus RNA. J Virol. 1977 Apr;22(1):235–237. doi: 10.1128/jvi.22.1.235-237.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Leung W. C., Leung M. F., Rawls W. E. Distinctive RNA transcriptase, polyadenylic acid polymerase, and polyuridylic acid polymerase activities associated with Pichinde virus. J Virol. 1979 Apr;30(1):98–107. doi: 10.1128/jvi.30.1.98-107.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Leung W. C., Ramsingh A., Dimock K., Rawls W. E., Petrovich J., Leung M. Pichinde virus L and S RNAs contain unique sequences. J Virol. 1981 Jan;37(1):48–54. doi: 10.1128/jvi.37.1.48-54.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Morrison T. G. Site of synthesis of membrane and nonmembrane proteins of vesicular stomatitis virus. J Biol Chem. 1975 Sep 10;250(17):6955–6962. [PubMed] [Google Scholar]
  12. Paul M., Goldsmith M. R., Hunsley J. R., Kafatos F. C. Specific protein synthesis in cellular differentiation. Production of eggshell proteins by silkmoth follicular cells. J Cell Biol. 1972 Dec;55(3):653–680. doi: 10.1083/jcb.55.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ramos B. A., Courtney R. J., Rawls W. E. Structural proteins of Pichinde virus. J Virol. 1972 Oct;10(4):661–667. doi: 10.1128/jvi.10.4.661-667.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ramsingh A. I., Dimock K., Rawls W. E., Leung W. C. Size estimation of Pichinde virus RNA by gel electrophoresis under denaturing conditions. Intervirology. 1980;14(1):31–36. doi: 10.1159/000149159. [DOI] [PubMed] [Google Scholar]
  15. Tkacz J. S., Lampen O. Tunicamycin inhibition of polyisoprenyl N-acetylglucosaminyl pyrophosphate formation in calf-liver microsomes. Biochem Biophys Res Commun. 1975 Jul 8;65(1):248–257. doi: 10.1016/s0006-291x(75)80086-6. [DOI] [PubMed] [Google Scholar]
  16. Vezza A. C., Gard G. P., Compans R. W., Bishop D. H. Structural components of the arenavirus Pichinde. J Virol. 1977 Sep;23(3):776–786. doi: 10.1128/jvi.23.3.776-786.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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