Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1995 Jul;69(7):4519–4523. doi: 10.1128/jvi.69.7.4519-4523.1995

A molecular determinant of human immunodeficiency virus particle assembly located in matrix antigen p17.

Y Morikawa 1, T Kishi 1, W H Zhang 1, M V Nermut 1, D J Hockley 1, I M Jones 1
PMCID: PMC189197  PMID: 7769715

Abstract

We report single-point mutations that are located in the matrix protein domain of the gag gene of human immunodeficiency virus type 1 and that prevent Gag particle formation. We show that mutations of p17 that abolish human immunodeficiency virus particle assembly also prevent the dimerization of p17 protein, as measured directly by a protein-protein binding assay. In the three-dimensional structure of p17, mutations that abolish dimerization are located in a single alpha helix that forms part of a fingerlike projection from one side of the molecule. Peptides derived from this region of p17 also reduce the level of p17 dimer when they are added to p17-expressing cells and compete for p17 self-association when present in protein-protein binding assays. We propose that the dimerization of the Gag precursor that occurs by the interdigitation of alpha helices on adjacent matrix molecules is a key stage in virion assembly and that the prevention of such an interaction is the molecular basis of particle misassembly.

Full Text

The Full Text of this article is available as a PDF (855.0 KB).

Selected References

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

  1. 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]
  2. Bukrinsky M. I., Haggerty S., Dempsey M. P., Sharova N., Adzhubel A., Spitz L., Lewis P., Goldfarb D., Emerman M., Stevenson M. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature. 1993 Oct 14;365(6447):666–669. doi: 10.1038/365666a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chazal N., Carrière C., Gay B., Boulanger P. Phenotypic characterization of insertion mutants of the human immunodeficiency virus type 1 Gag precursor expressed in recombinant baculovirus-infected cells. J Virol. 1994 Jan;68(1):111–122. doi: 10.1128/jvi.68.1.111-122.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chazal N., Gay B., Carrière C., Tournier J., Boulanger P. Human immunodeficiency virus type 1 MA deletion mutants expressed in baculovirus-infected cells: cis and trans effects on the Gag precursor assembly pathway. J Virol. 1995 Jan;69(1):365–375. doi: 10.1128/jvi.69.1.365-375.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Delchambre M., Gheysen D., Thines D., Thiriart C., Jacobs E., Verdin E., Horth M., Burny A., Bex F. The GAG precursor of simian immunodeficiency virus assembles into virus-like particles. EMBO J. 1989 Sep;8(9):2653–2660. doi: 10.1002/j.1460-2075.1989.tb08405.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deminie C. A., Emerman M. Incorporation of human immunodeficiency virus type 1 Gag proteins into murine leukemia virus virions. J Virol. 1993 Nov;67(11):6499–6506. doi: 10.1128/jvi.67.11.6499-6506.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ehrlich L. S., Agresta B. E., Carter C. A. Assembly of recombinant human immunodeficiency virus type 1 capsid protein in vitro. J Virol. 1992 Aug;66(8):4874–4883. doi: 10.1128/jvi.66.8.4874-4883.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Freed E. O., Orenstein J. M., Buckler-White A. J., Martin M. A. Single amino acid changes in the human immunodeficiency virus type 1 matrix protein block virus particle production. J Virol. 1994 Aug;68(8):5311–5320. doi: 10.1128/jvi.68.8.5311-5320.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fäcke M., Janetzko A., Shoeman R. L., Kräusslich H. G. A large deletion in the matrix domain of the human immunodeficiency virus gag gene redirects virus particle assembly from the plasma membrane to the endoplasmic reticulum. J Virol. 1993 Aug;67(8):4972–4980. doi: 10.1128/jvi.67.8.4972-4980.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gelderblom H. R., Hausmann E. H., Ozel M., Pauli G., Koch M. A. Fine structure of human immunodeficiency virus (HIV) and immunolocalization of structural proteins. Virology. 1987 Jan;156(1):171–176. doi: 10.1016/0042-6822(87)90449-1. [DOI] [PubMed] [Google Scholar]
  11. Gheysen D., Jacobs E., de Foresta F., Thiriart C., Francotte M., Thines D., De Wilde M. Assembly and release of HIV-1 precursor Pr55gag virus-like particles from recombinant baculovirus-infected insect cells. Cell. 1989 Oct 6;59(1):103–112. doi: 10.1016/0092-8674(89)90873-8. [DOI] [PubMed] [Google Scholar]
  12. Gilmour J. E., Senior J. M., Burns N. R., Esnouf M. P., Gull K., Kingsman S. M., Kingsman A. J., Adams S. E. A novel method for the purification of HIV-1 p24 protein from hybrid Ty virus-like particles (Ty-VLPs). AIDS. 1989 Nov;3(11):717–723. doi: 10.1097/00002030-198911000-00005. [DOI] [PubMed] [Google Scholar]
  13. Grässer F. A., Sauder C., Haiss P., Hille A., König S., Göttel S., Kremmer E., Leinenbach H. P., Zeppezauer M., Mueller-Lantzsch N. Immunological detection of proteins associated with the Epstein-Barr virus nuclear antigen 2A. Virology. 1993 Aug;195(2):550–560. doi: 10.1006/viro.1993.1406. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Hockley D. J., Wood R. D., Jacobs J. P., Garrett A. J. Electron microscopy of human immunodeficiency virus. J Gen Virol. 1988 Oct;69(Pt 10):2455–2469. doi: 10.1099/0022-1317-69-10-2455. [DOI] [PubMed] [Google Scholar]
  16. Homann H. E., Willenbrink W., Buchholz C. J., Neubert W. J. Sendai virus protein-protein interactions studied by a protein-blotting protein-overlay technique: mapping of domains on NP protein required for binding to P protein. J Virol. 1991 Mar;65(3):1304–1309. doi: 10.1128/jvi.65.3.1304-1309.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hong S. S., Boulanger P. Assembly-defective point mutants of the human immunodeficiency virus type 1 Gag precursor phenotypically expressed in recombinant baculovirus-infected cells. J Virol. 1993 May;67(5):2787–2798. doi: 10.1128/jvi.67.5.2787-2798.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoshikawa N., Kojima A., Yasuda A., Takayashiki E., Masuko S., Chiba J., Sata T., Kurata T. Role of the gag and pol genes of human immunodeficiency virus in the morphogenesis and maturation of retrovirus-like particles expressed by recombinant vaccinia virus: an ultrastructural study. J Gen Virol. 1991 Oct;72(Pt 10):2509–2517. doi: 10.1099/0022-1317-72-10-2509. [DOI] [PubMed] [Google Scholar]
  19. Hu S. L., Travis B. M., Garrigues J., Zarling J. M., Sridhar P., Dykers T., Eichberg J. W., Alpers C. Processing, assembly, and immunogenicity of human immunodeficiency virus core antigens expressed by recombinant vaccinia virus. Virology. 1990 Nov;179(1):321–329. doi: 10.1016/0042-6822(90)90300-g. [DOI] [PubMed] [Google Scholar]
  20. Jowett J. B., Hockley D. J., Nermut M. V., Jones I. M. Distinct signals in human immunodeficiency virus type 1 Pr55 necessary for RNA binding and particle formation. J Gen Virol. 1992 Dec;73(Pt 12):3079–3086. doi: 10.1099/0022-1317-73-12-3079. [DOI] [PubMed] [Google Scholar]
  21. Karacostas V., Nagashima K., Gonda M. A., Moss B. Human immunodeficiency virus-like particles produced by a vaccinia virus expression vector. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8964–8967. doi: 10.1073/pnas.86.22.8964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kitts P. A., Ayres M. D., Possee R. D. Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Res. 1990 Oct 11;18(19):5667–5672. doi: 10.1093/nar/18.19.5667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  24. Livingstone C., Jones I. Baculovirus expression vectors with single strand capability. Nucleic Acids Res. 1989 Mar 25;17(6):2366–2366. doi: 10.1093/nar/17.6.2366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Luban J., Lee C., Goff S. P. Effect of linker insertion mutations in the human immunodeficiency virus type 1 gag gene on activation of viral protease expressed in bacteria. J Virol. 1993 Jun;67(6):3630–3634. doi: 10.1128/jvi.67.6.3630-3634.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Massiah M. A., Starich M. R., Paschall C., Summers M. F., Christensen A. M., Sundquist W. I. Three-dimensional structure of the human immunodeficiency virus type 1 matrix protein. J Mol Biol. 1994 Nov 25;244(2):198–223. doi: 10.1006/jmbi.1994.1719. [DOI] [PubMed] [Google Scholar]
  27. Matthews S., Barlow P., Boyd J., Barton G., Russell R., Mills H., Cunningham M., Meyers N., Burns N., Clark N. Structural similarity between the p17 matrix protein of HIV-1 and interferon-gamma. Nature. 1994 Aug 25;370(6491):666–668. doi: 10.1038/370666a0. [DOI] [PubMed] [Google Scholar]
  28. Mergener K., Fäcke M., Welker R., Brinkmann V., Gelderblom H. R., Kräusslich H. G. Analysis of HIV particle formation using transient expression of subviral constructs in mammalian cells. Virology. 1992 Jan;186(1):25–39. doi: 10.1016/0042-6822(92)90058-w. [DOI] [PubMed] [Google Scholar]
  29. Mills H. R., Jones I. M. Expression and purification of p24, the core protein of HIV, using a baculovirus-insect cell expression system. AIDS. 1990 Nov;4(11):1125–1131. doi: 10.1097/00002030-199011000-00011. [DOI] [PubMed] [Google Scholar]
  30. Nermut M. V., Grief C., Hashmi S., Hockley D. J. Further evidence of icosahedral symmetry in human and simian immunodeficiency virus. AIDS Res Hum Retroviruses. 1993 Oct;9(10):929–938. doi: 10.1089/aid.1993.9.929. [DOI] [PubMed] [Google Scholar]
  31. Nermut M. V., Hockley D. J., Jowett J. B., Jones I. M., Garreau M., Thomas D. Fullerene-like organization of HIV gag-protein shell in virus-like particles produced by recombinant baculovirus. Virology. 1994 Jan;198(1):288–296. doi: 10.1006/viro.1994.1032. [DOI] [PubMed] [Google Scholar]
  32. Niedrig M., Gelderblom H. R., Pauli G., März J., Bickhard H., Wolf H., Modrow S. Inhibition of infectious human immunodeficiency virus type 1 particle formation by Gag protein-derived peptides. J Gen Virol. 1994 Jun;75(Pt 6):1469–1474. doi: 10.1099/0022-1317-75-6-1469. [DOI] [PubMed] [Google Scholar]
  33. Overton H. A., Fujii Y., Price I. R., Jones I. M. The protease and gag gene products of the human immunodeficiency virus: authentic cleavage and post-translational modification in an insect cell expression system. Virology. 1989 May;170(1):107–116. doi: 10.1016/0042-6822(89)90357-7. [DOI] [PubMed] [Google Scholar]
  34. Royer M., Cerutti M., Gay B., Hong S. S., Devauchelle G., Boulanger P. Functional domains of HIV-1 gag-polyprotein expressed in baculovirus-infected cells. Virology. 1991 Sep;184(1):417–422. doi: 10.1016/0042-6822(91)90861-5. [DOI] [PubMed] [Google Scholar]
  35. Shioda T., Shibuta H. Production of human immunodeficiency virus (HIV)-like particles from cells infected with recombinant vaccinia viruses carrying the gag gene of HIV. Virology. 1990 Mar;175(1):139–148. doi: 10.1016/0042-6822(90)90194-v. [DOI] [PubMed] [Google Scholar]
  36. Smith A. J., Cho M. I., Hammarskjöld M. L., Rekosh D. Human immunodeficiency virus type 1 Pr55gag and Pr160gag-pol expressed from a simian virus 40 late replacement vector are efficiently processed and assembled into viruslike particles. J Virol. 1990 Jun;64(6):2743–2750. doi: 10.1128/jvi.64.6.2743-2750.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  38. Trono D., Feinberg M. B., Baltimore D. HIV-1 Gag mutants can dominantly interfere with the replication of the wild-type virus. Cell. 1989 Oct 6;59(1):113–120. doi: 10.1016/0092-8674(89)90874-x. [DOI] [PubMed] [Google Scholar]
  39. Veronese F. D., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Biochemical and immunological analysis of human immunodeficiency virus gag gene products p17 and p24. J Virol. 1988 Mar;62(3):795–801. doi: 10.1128/jvi.62.3.795-801.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang C. T., Barklis E. Assembly, processing, and infectivity of human immunodeficiency virus type 1 gag mutants. J Virol. 1993 Jul;67(7):4264–4273. doi: 10.1128/jvi.67.7.4264-4273.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Weiss S., König B., Morikawa Y., Jones I. Recombinant HIV-1 nucleocapsid protein p15 produced as a fusion protein with glutathione S-transferase in Escherichia coli mediates dimerization and enhances reverse transcription of retroviral RNA. Gene. 1992 Nov 16;121(2):203–212. doi: 10.1016/0378-1119(92)90123-7. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Yu X., Yuan X., Matsuda Z., Lee T. H., Essex M. The matrix protein of human immunodeficiency virus type 1 is required for incorporation of viral envelope protein into mature virions. J Virol. 1992 Aug;66(8):4966–4971. doi: 10.1128/jvi.66.8.4966-4971.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yuan X., Yu X., Lee T. H., Essex M. Mutations in the N-terminal region of human immunodeficiency virus type 1 matrix protein block intracellular transport of the Gag precursor. J Virol. 1993 Nov;67(11):6387–6394. doi: 10.1128/jvi.67.11.6387-6394.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Zhoa Y., Jones I. M., Hockley D. J., Nermut M. V., Roy P. Complementation of human immunodeficiency virus (HIV-1) gag particle formation. Virology. 1994 Mar;199(2):403–408. doi: 10.1006/viro.1994.1138. [DOI] [PubMed] [Google Scholar]
  46. von Poblotzki A., Wagner R., Niedrig M., Wanner G., Wolf H., Modrow S. Identification of a region in the Pr55gag-polyprotein essential for HIV-1 particle formation. Virology. 1993 Apr;193(2):981–985. doi: 10.1006/viro.1993.1210. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES