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. 1996 Aug;70(8):5695–5700. doi: 10.1128/jvi.70.8.5695-5700.1996

Fine mapping and characterization of the Rous sarcoma virus Pr76gag late assembly domain.

Y Xiang 1, C E Cameron 1, J W Wills 1, J Leis 1
PMCID: PMC190537  PMID: 8764091

Abstract

The p2 region of the Rous sarcoma virus (RSV) Gag polyprotein contains an assembly domain, which is required late in replication for efficient budding of virus-like particles from cells (J. W. Wills, C. E. Cameron, C. B. Wilson, Y. Xiang, R. P. Bennett, and J. Leis, J. Virol. 68:6605-6618, 1994). This domain, referred to as the L domain, was previously mapped to the 11 amino acids of p2b. Through the analysis of a series of deletion and substitution mutations, the L domain has now been fine mapped to a highly conserved amino acid sequence, PPPPYV of p2b. Sequences flanking PPPPYV motif can be deleted without any effect on budding. Defects caused by L-domain deletions can be rescued by placing a wild-type copy of the sequence at several other positions in RSV Gag. A proline-rich P(S/T)APP motif is found in many retroviral Gag polyproteins; the motif found in the p6 region of human immunodeficiency virus type 1 has been implicated in late functions of the virus. Substitution of the RSV L domain with this motif in a 10-amino-acid sequence derived from visna leukemia virus results in wild-type release of virus particles from cells. In contrast, the slightly different sequences from Gibbon ape leukemia virus, Moloney leukemia virus, PSAPP alone, or a proline-rich SH3 binding sequence do not efficiently rescue RSV L-domain mutations.

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Selected References

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  1. Aiyar A., Xiang Y., Leis J. Site-directed mutagenesis using overlap extension PCR. Methods Mol Biol. 1996;57:177–191. doi: 10.1385/0-89603-332-5:177. [DOI] [PubMed] [Google Scholar]
  2. Bennett R. P., Nelle T. D., Wills J. W. Functional chimeras of the Rous sarcoma virus and human immunodeficiency virus gag proteins. J Virol. 1993 Nov;67(11):6487–6498. doi: 10.1128/jvi.67.11.6487-6498.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett R. P., Rhee S., Craven R. C., Hunter E., Wills J. W. Amino acids encoded downstream of gag are not required by Rous sarcoma virus protease during gag-mediated assembly. J Virol. 1991 Jan;65(1):272–280. doi: 10.1128/jvi.65.1.272-280.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowles N., Bonnet D., Mulhauser F., Spahr P. F. Site-directed mutagenesis of the P2 region of the Rous sarcoma virus gag gene: effects on Gag polyprotein processing. Virology. 1994 Aug 15;203(1):20–28. doi: 10.1006/viro.1994.1450. [DOI] [PubMed] [Google Scholar]
  5. Cameron C. E., Grinde B., Jacques P., Jentoft J., Leis J., Wlodawer A., Weber I. T. Comparison of the substrate-binding pockets of the Rous sarcoma virus and human immunodeficiency virus type 1 proteases. J Biol Chem. 1993 Jun 5;268(16):11711–11720. [PubMed] [Google Scholar]
  6. Cameron C. E., Grinde B., Jentoft J., Leis J., Weber I. T., Copeland T. D., Wlodawer A. Mechanism of inhibition of the retroviral protease by a Rous sarcoma virus peptide substrate representing the cleavage site between the gag p2 and p10 proteins. J Biol Chem. 1992 Nov 25;267(33):23735–23741. [PubMed] [Google Scholar]
  7. Chen H. I., Sudol M. The WW domain of Yes-associated protein binds a proline-rich ligand that differs from the consensus established for Src homology 3-binding modules. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7819–7823. doi: 10.1073/pnas.92.17.7819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Craven R. C., Bennett R. P., Wills J. W. Role of the avian retroviral protease in the activation of reverse transcriptase during virion assembly. J Virol. 1991 Nov;65(11):6205–6217. doi: 10.1128/jvi.65.11.6205-6217.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Craven R. C., Leure-duPree A. E., Erdie C. R., Wilson C. B., Wills J. W. Necessity of the spacer peptide between CA and NC in the Rous sarcoma virus gag protein. J Virol. 1993 Oct;67(10):6246–6252. doi: 10.1128/jvi.67.10.6246-6252.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Franke E. K., Yuan H. E., Luban J. Specific incorporation of cyclophilin A into HIV-1 virions. Nature. 1994 Nov 24;372(6504):359–362. doi: 10.1038/372359a0. [DOI] [PubMed] [Google Scholar]
  11. Grinde B., Cameron C. E., Leis J., Weber I. T., Wlodawer A., Burstein H., Bizub D., Skalka A. M. Mutations that alter the activity of the Rous sarcoma virus protease. J Biol Chem. 1992 May 15;267(14):9481–9490. [PubMed] [Google Scholar]
  12. Göttlinger H. G., Dorfman T., Sodroski J. G., Haseltine W. A. Effect of mutations affecting the p6 gag protein on human immunodeficiency virus particle release. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3195–3199. doi: 10.1073/pnas.88.8.3195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Huang M., Orenstein J. M., Martin M. A., Freed E. O. p6Gag is required for particle production from full-length human immunodeficiency virus type 1 molecular clones expressing protease. J Virol. 1995 Nov;69(11):6810–6818. doi: 10.1128/jvi.69.11.6810-6818.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leis J., Phillips N., Fu X., Tuazon P. T., Traugh J. A. Phosphorylation of avian retrovirus matrix protein by Ca2+/phospholipid-dependent protein kinase. Eur J Biochem. 1989 Feb 1;179(2):415–422. doi: 10.1111/j.1432-1033.1989.tb14569.x. [DOI] [PubMed] [Google Scholar]
  15. Parent L. J., Bennett R. P., Craven R. C., Nelle T. D., Krishna N. K., Bowzard J. B., Wilson C. B., Puffer B. A., Montelaro R. C., Wills J. W. Positionally independent and exchangeable late budding functions of the Rous sarcoma virus and human immunodeficiency virus Gag proteins. J Virol. 1995 Sep;69(9):5455–5460. doi: 10.1128/jvi.69.9.5455-5460.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Parent L. J., Wilson C. B., Resh M. D., Wills J. W. Evidence for a second function of the MA sequence in the Rous sarcoma virus Gag protein. J Virol. 1996 Feb;70(2):1016–1026. doi: 10.1128/jvi.70.2.1016-1026.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pepinsky R. B., Papayannopoulos I. A., Chow E. P., Krishna N. K., Craven R. C., Vogt V. M. Differential proteolytic processing leads to multiple forms of the CA protein in avian sarcoma and leukemia viruses. J Virol. 1995 Oct;69(10):6430–6438. doi: 10.1128/jvi.69.10.6430-6438.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Skalka A. M. Retroviral proteases: first glimpses at the anatomy of a processing machine. Cell. 1989 Mar 24;56(6):911–913. doi: 10.1016/0092-8674(89)90621-1. [DOI] [PubMed] [Google Scholar]
  19. Sudol M., Bork P., Einbond A., Kastury K., Druck T., Negrini M., Huebner K., Lehman D. Characterization of the mammalian YAP (Yes-associated protein) gene and its role in defining a novel protein module, the WW domain. J Biol Chem. 1995 Jun 16;270(24):14733–14741. doi: 10.1074/jbc.270.24.14733. [DOI] [PubMed] [Google Scholar]
  20. Sudol M., Chen H. I., Bougeret C., Einbond A., Bork P. Characterization of a novel protein-binding module--the WW domain. FEBS Lett. 1995 Aug 1;369(1):67–71. doi: 10.1016/0014-5793(95)00550-s. [DOI] [PubMed] [Google Scholar]
  21. Weldon R. A., Jr, Erdie C. R., Oliver M. G., Wills J. W. Incorporation of chimeric gag protein into retroviral particles. J Virol. 1990 Sep;64(9):4169–4179. doi: 10.1128/jvi.64.9.4169-4179.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weldon R. A., Jr, Wills J. W. Characterization of a small (25-kilodalton) derivative of the Rous sarcoma virus Gag protein competent for particle release. J Virol. 1993 Sep;67(9):5550–5561. doi: 10.1128/jvi.67.9.5550-5561.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wills J. W., Cameron C. E., Wilson C. B., Xiang Y., Bennett R. P., Leis J. An assembly domain of the Rous sarcoma virus Gag protein required late in budding. J Virol. 1994 Oct;68(10):6605–6618. doi: 10.1128/jvi.68.10.6605-6618.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wills J. W., Craven R. C., Achacoso J. A. Creation and expression of myristylated forms of Rous sarcoma virus gag protein in mammalian cells. J Virol. 1989 Oct;63(10):4331–4343. doi: 10.1128/jvi.63.10.4331-4343.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wills J. W., Craven R. C. Form, function, and use of retroviral gag proteins. AIDS. 1991 Jun;5(6):639–654. doi: 10.1097/00002030-199106000-00002. [DOI] [PubMed] [Google Scholar]
  26. Wills J. W., Craven R. C., Weldon R. A., Jr, Nelle T. D., Erdie C. R. Suppression of retroviral MA deletions by the amino-terminal membrane-binding domain of p60src. J Virol. 1991 Jul;65(7):3804–3812. doi: 10.1128/jvi.65.7.3804-3812.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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