Abstract
The internal structural proteins of avian sarcoma and leukemia viruses are derived from a precursor polypeptide that is the product of the viral gag gene. The N-terminal domain of the precursor gives rise to p19, a protein that interacts with the lipid envelope of the virus and that may also interact with viral RNA. The C terminus of p19 from the Prague C strain of Rous sarcoma virus was previously assigned to a tyrosine residue 175 amino acids from the N terminus. We have used metabolic labeling and carboxypeptidase digestion to show that the C terminus of p19 is actually tyrosine 155. This implies the existence of a sixth gag protein 22 amino acids in length and located between p19 and p10 on the gag precursor. The p19 species of some recombinant avian sarcoma viruses and of the defective endogenous virus derived from the ev-1 locus migrate on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as if they were about 4,000 daltons smaller than p19. We have elucidated the structure of these forms, called p19 beta, by analysis of the proteins and determination of the DNA sequence of the p19 region of the gag gene from ev-1 and ev-2. Esterification of carboxyl groups completely suppressed the differences in migration of p19 and p19 beta. Peptide mapping showed the altered mobility to be determined by sequences in the C-terminal cyanogen bromide fragment of the proteins. We conclude from the DNA sequence that a single glutamate-lysine alteration is responsible for the altered electrophoretic mobility.
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- 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]
- Bizub D., Katz R. A., Skalka A. M. Nucleotide sequence of noncoding regions in Rous-associated virus-2: comparisons delineate conserved regions important in replication and oncogenesis. J Virol. 1984 Feb;49(2):557–565. doi: 10.1128/jvi.49.2.557-565.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coffin J. M., Tsichlis P. N., Conklin K. F., Senior A., Robinson H. L. Genomes of endogenous and exogenous avian retroviruses. Virology. 1983 Apr 15;126(1):51–72. doi: 10.1016/0042-6822(83)90461-0. [DOI] [PubMed] [Google Scholar]
- Conklin K. F., Coffin J. M., Robinson H. L., Groudine M., Eisenman R. Role of methylation in the induced and spontaneous expression of the avian endogenous virus ev-1: DNA structure and gene products. Mol Cell Biol. 1982 Jun;2(6):638–652. doi: 10.1128/mcb.2.6.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cullen B. R., Skalka A. M., Ju G. Endogenous avian retroviruses contain deficient promoter and leader sequences. Proc Natl Acad Sci U S A. 1983 May;80(10):2946–2950. doi: 10.1073/pnas.80.10.2946. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eisenman R. N., Vogt V. M. The biosynthesis of oncovirus proteins. Biochim Biophys Acta. 1978 Apr 6;473(3-4):187–239. doi: 10.1016/0304-419x(78)90014-8. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- Johnson S. P., Veigl M., Vanaman T., Leis J. Cyanogen bromide digestion of the avian myeloblastosis virus pp19 protein: isolation of an amino-terminal peptide that binds to viral RNA. J Virol. 1983 Feb;45(2):876–881. doi: 10.1128/jvi.45.2.876-881.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kitamura N., Kitamura A., Toyoshima K., Hirayama Y., Yoshida M. Avian sarcoma virus Y73 genome sequence and structural similarity of its transforming gene product to that of Rous sarcoma virus. Nature. 1982 May 20;297(5863):205–208. doi: 10.1038/297205a0. [DOI] [PubMed] [Google Scholar]
- Leis J. P., McGinnis J., Green R. W. Rous sarcoma virus p19 binds to specific double-stranded regions of viral RNA: effect of p19 on cleavage of viral RNA by RNase III. Virology. 1978 Jan;84(1):87–98. doi: 10.1016/0042-6822(78)90220-9. [DOI] [PubMed] [Google Scholar]
- Linial M., Medeiros E., Hayward W. S. An avian oncovirus mutant (SE 21Q1b) deficient in genomic RNA: biological and biochemical characterization. Cell. 1978 Dec;15(4):1371–1381. doi: 10.1016/0092-8674(78)90062-4. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Palmiter R. D., Gagnon J., Vogt V. M., Ripley S., Eisenman R. N. The NH2-terminal sequence of the avian oncovirus gag precursor polyprotein (Pr76gag). Virology. 1978 Dec;91(2):423–433. doi: 10.1016/0042-6822(78)90388-4. [DOI] [PubMed] [Google Scholar]
- Pepinsky R. B. Localization of lipid-protein and protein-protein interactions within the murine retrovirus gag precursor by a novel peptide-mapping technique. J Biol Chem. 1983 Sep 25;258(18):11229–11235. [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Reddy E. P., Reynolds R. K., Watson D. K., Schultz R. A., Lautenberger J., Papas T. S. Nucleotide sequence analysis of the proviral genome of avian myelocytomatosis virus (MC29). Proc Natl Acad Sci U S A. 1983 May;80(9):2500–2504. doi: 10.1073/pnas.80.9.2500. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Schwartz W. E., Smith P. K., Royer G. P. N-(beta-Iodoethyl)trifluoroacetamide: a new reagent for the aminoethylation of thio groups in proteins. Anal Biochem. 1980 Jul 15;106(1):43–48. doi: 10.1016/0003-2697(80)90116-5. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Shibuya M., Hanafusa H. Nucleotide sequence of Fujinami sarcoma virus: evolutionary relationship of its transforming gene with transforming genes of other sarcoma viruses. Cell. 1982 Oct;30(3):787–795. doi: 10.1016/0092-8674(82)90283-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Swanstrom R., Varmus H. E., Bishop J. M. Nucleotide sequence of the 5' noncoding region and part of the gag gene of Rous sarcoma virus. J Virol. 1982 Feb;41(2):535–541. doi: 10.1128/jvi.41.2.535-541.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tung J. S., Knight C. A. Relative importance of some factors affecting the electrophoretic migration of proteins in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1972 Jul;48(1):153–163. doi: 10.1016/0003-2697(72)90179-0. [DOI] [PubMed] [Google Scholar]
- Versteegen R. J., Copeland T. D., Oroszlan S. Complete amino acid sequence of the group-specific antigen gene-encoded phosphorylated proteins of mouse leukemia viruses. J Biol Chem. 1982 Mar 25;257(6):3007–3013. [PubMed] [Google Scholar]
- Vogt V. M., Eisenman R., Diggelmann H. Generation of avian myeloblastosis virus structural proteins by proteolytic cleavage of a precursor polypeptide. J Mol Biol. 1975 Aug 15;96(3):471–493. doi: 10.1016/0022-2836(75)90174-6. [DOI] [PubMed] [Google Scholar]
- 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]



