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. 1986 Apr;58(1):50–58. doi: 10.1128/jvi.58.1.50-58.1986

Structure and processing of the p2 region of avian sarcoma and leukemia virus gag precursor polyproteins.

R B Pepinsky, R J Mattaliano, V M Vogt
PMCID: PMC252875  PMID: 3005658

Abstract

We have purified two low-molecular-weight polypeptides from the Prague C strain of Rous sarcoma virus and have identified these as products of the gag precursor Pr76 by protein sequencing and by amino acid analysis. Both polypeptides are derived from a stretch of 22 amino acids within Pr76 that separates p19 and p10. We refer to this region as p2. Together the two cleavage products form the entire p2 region. The junctions of p19 with the amino-terminal fragment of p2 and of p10 with the carboxy-terminal fragment of p2 define two new processing sites within the gag precursor, Tyr-155-His-156 and Gly-177-Ser-178. Both polypeptides are major cleavage products of Pr76 that occur in Prague C Rous sarcoma virus at an estimated 1,000 copies per virion. They also are prominent components of avian myeloblastosis virus. The combination of gel filtration and reverse-phase high-pressure liquid chromatography, which was used for the isolation of the two fragments of p2, resolved over a dozen other low-molecular-weight polypeptides from avian sarcoma and leukemia viruses that previously were undetected. This technique thus should serve as a useful procedure for further characterization of viral components.

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Selected References

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  1. Bhown A. S., Bennett J. C., Hunter E. Alignment of the peptides derived from acid-catalyzed cleavage of an aspartylprolyl bond in the major internal structural polypeptide of avian retroviruses. J Biol Chem. 1980 Jul 25;255(14):6962–6965. [PubMed] [Google Scholar]
  2. Bhown A. S., Bennett J. C., Mole J. E., Hunter E. Purification and characterization of the gag gene products of avian-type C retroviruses by high-pressure liquid chromatography. Anal Biochem. 1981 Mar 15;112(1):128–134. doi: 10.1016/0003-2697(81)90269-4. [DOI] [PubMed] [Google Scholar]
  3. Bolognesi D. P., Ishizaki R., Hüper G., Vanaman T. C., Smith R. E. Immunological properties of avian oncornavirus polypeptides. Virology. 1975 Apr;64(2):349–357. doi: 10.1016/0042-6822(75)90111-7. [DOI] [PubMed] [Google Scholar]
  4. Bolognesi D. P., Luftig R., Shaper J. H. Localization of RNA tumor virus polypeptides. I. Isolation of further virus substructures. Virology. 1973 Dec;56(2):549–564. doi: 10.1016/0042-6822(73)90057-3. [DOI] [PubMed] [Google Scholar]
  5. Broome S., Gilbert W. Rous sarcoma virus encodes a transcriptional activator. Cell. 1985 Mar;40(3):537–546. doi: 10.1016/0092-8674(85)90202-8. [DOI] [PubMed] [Google Scholar]
  6. Davis N. L., Rueckert R. R. Properties of a ribonucleoprotein particle isolated from Nonidet P-40-treated Rous sarcoma virus. J Virol. 1972 Nov;10(5):1010–1020. doi: 10.1128/jvi.10.5.1010-1020.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dittmar K. J., Moelling K. Biochemical properties of p15-associated protease in an avian RNA tumor virus. J Virol. 1978 Oct;28(1):106–118. doi: 10.1128/jvi.28.1.106-118.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fletcher P., Nowinski R. C., Tress E., Fleissner E. Chromatographic separation and antigenic analysis of proteins of the oncornaviruses. III. Avian viral proteins with group-specific antigenicity. Virology. 1975 Apr;64(2):358–366. doi: 10.1016/0042-6822(75)90112-9. [DOI] [PubMed] [Google Scholar]
  9. Gebhardt A., Bosch J. V., Ziemiecki A., Friis R. R. Rous sarcoma virus p19 and gp35 can be chemically crosslinked to high molecular weight complexes. An insight into virus assembly. J Mol Biol. 1984 Apr 5;174(2):297–317. doi: 10.1016/0022-2836(84)90340-1. [DOI] [PubMed] [Google Scholar]
  10. Hampe A., Laprevotte I., Galibert F., Fedele L. A., Sherr C. J. Nucleotide sequences of feline retroviral oncogenes (v-fes) provide evidence for a family of tyrosine-specific protein kinase genes. Cell. 1982 Oct;30(3):775–785. doi: 10.1016/0092-8674(82)90282-3. [DOI] [PubMed] [Google Scholar]
  11. Herman A. C., Green R. W., Bolognesi D. P., Vanaman T. C. Comparative chemical properties of avian oncornavirus polypeptides. Virology. 1975 Apr;64(2):339–348. doi: 10.1016/0042-6822(75)90110-5. [DOI] [PubMed] [Google Scholar]
  12. Hunkapiller M. W., Hood L. E. Analysis of phenylthiohydantoins by ultrasensitive gradient high-performance liquid chromatography. Methods Enzymol. 1983;91:486–493. doi: 10.1016/s0076-6879(83)91045-5. [DOI] [PubMed] [Google Scholar]
  13. Hunter E., Bennett J. C., Bhown A., Pepinsky R. B., Vogt V. M. Amino-terminal amino acid sequence of p10, the fifth major gag polypeptide of avian sarcoma and leukemia viruses. J Virol. 1983 Feb;45(2):885–888. doi: 10.1128/jvi.45.2.885-888.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. O'Rear J. J., Temin H. M. Spontaneous changes in nucleotide sequence in proviruses of spleen necrosis virus, an avian retrovirus. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1230–1234. doi: 10.1073/pnas.79.4.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pepinsky R. B., Vogt V. M. Fine-structure analyses of lipid-protein and protein-protein interactions of gag protein p19 of the avian sarcoma and leukemia viruses by cyanogen bromide mapping. J Virol. 1984 Oct;52(1):145–153. doi: 10.1128/jvi.52.1.145-153.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pepinsky R. B., Vogt V. M. Identification of retrovirus matrix proteins by lipid-protein cross-linking. J Mol Biol. 1979 Jul 15;131(4):819–837. doi: 10.1016/0022-2836(79)90203-1. [DOI] [PubMed] [Google Scholar]
  17. Pepinsky R. B., Vogt V. M. Purification and properties of a fifth major viral gag protein from avian sarcoma and leukemia viruses. J Virol. 1983 Feb;45(2):648–658. doi: 10.1128/jvi.45.2.648-658.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sauer R. T., Allen D. W., Niall H. D. Amino acid sequence of p15 from avian myeloblastosis virus complex. Biochemistry. 1981 Jun 23;20(13):3784–3791. doi: 10.1021/bi00516a018. [DOI] [PubMed] [Google Scholar]
  19. Schwartz D. E., Tizard R., Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. doi: 10.1016/0092-8674(83)90071-5. [DOI] [PubMed] [Google Scholar]
  20. Seiki M., Hattori S., Hirayama Y., Yoshida M. Human adult T-cell leukemia virus: complete nucleotide sequence of the provirus genome integrated in leukemia cell DNA. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3618–3622. doi: 10.1073/pnas.80.12.3618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shaikh R., Linial M., Brown S., Sen A., Eisenman R. Recombinant avian oncoviruses. II. Alterations in the gag proteins and evidence for intragenic recombination. Virology. 1979 Jan 30;92(2):463–481. doi: 10.1016/0042-6822(79)90150-8. [DOI] [PubMed] [Google Scholar]
  22. Shealy D. J., Mosser A. G., Rueckert R. R. Novel p19-related protein in Rous-associated virus type 61: implications for avian gag gene order. J Virol. 1980 May;34(2):431–437. doi: 10.1128/jvi.34.2.431-437.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  24. Stromberg K., Hurley N. E., Davis N. L., Rueckert R. R., Fleissner E. Structural studies of avian myeloblastosis virus: comparison of polypeptides in virion and core component by dodecyl sulfate-polyacrylamide gel electrophoresis. J Virol. 1974 Feb;13(2):513–528. doi: 10.1128/jvi.13.2.513-528.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vogt V. M., Pepinsky R. B., Southard L. E. Primary structure of p19 species of avian sarcoma and leukemia viruses. J Virol. 1985 Oct;56(1):31–39. doi: 10.1128/jvi.56.1.31-39.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vogt V. M., Wight A., Eisenman R. In vitro cleavage of avian retrovirus gag proteins by viral protease p15. Virology. 1979 Oct 15;98(1):154–167. doi: 10.1016/0042-6822(79)90534-8. [DOI] [PubMed] [Google Scholar]

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