Abstract
Interacting domains in human immunodeficiency virus type 1 (HIV-1) Gag precursor (Pr55gag) expressed in recombinant baculovirus-infected cells were investigated by three different methods: (i) trans rescue and coencapsidation of C-terminal deletion (amber) Gag mutants and Gag chimeras into retrovirus-like particles in complementation experiments with HIV-1 wild-type (WT) Pr55gag, (ii) Gag-Gag interactions in vitro in Gag ligand affinity blotting assays, and (iii) quantitative immunoelectron microscopy of retrovirus-like Gag particles, using a panel of monoclonal antibodies to probe the epitope accessibility of encapsidated HIV-1 WT Pr55gag. Four discrete regions, within residues 210 to 241, 277 to 306 (major homology region), and 307 to 333 in the capsid (CA) protein and residues 358 to 374 at the CA-spacer peptide 2 (sp2) junction, were found to have a significant influence on Gag trans-packaging efficiency. A fifth region, within residues 375 to 426, overlapping the sp2-nucleocapsid (NC) protein junction and most of the NC, seemed to be essential for stable inter-Gag binding in vitro. The coincidence of the two regions from 358 to 374 and 375 to 426 with an immunologically silent domain in WT Gag particles suggested that they could participate in direct Gag interactions.
Full Text
The Full Text of this article is available as a PDF (1.2 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Argos P. A possible homology between immunodeficiency virus p24 core protein and picornaviral VP2 coat protein: prediction of HIV p24 antigenic sites. EMBO J. 1989 Mar;8(3):779–785. doi: 10.1002/j.1460-2075.1989.tb03438.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Bryant M., Ratner L. Myristoylation-dependent replication and assembly of human immunodeficiency virus 1. Proc Natl Acad Sci U S A. 1990 Jan;87(2):523–527. doi: 10.1073/pnas.87.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Defer C., Belin M. T., Caillet-Boudin M. L., Boulanger P. Human adenovirus-host cell interactions: comparative study with members of subgroups B and C. J Virol. 1990 Aug;64(8):3661–3673. doi: 10.1128/jvi.64.8.3661-3673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorfman T., Luban J., Goff S. P., Haseltine W. A., Göttlinger H. G. Mapping of functionally important residues of a cysteine-histidine box in the human immunodeficiency virus type 1 nucleocapsid protein. J Virol. 1993 Oct;67(10):6159–6169. doi: 10.1128/jvi.67.10.6159-6169.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dorfman T., Mammano F., Haseltine W. A., Göttlinger H. G. Role of the matrix protein in the virion association of the human immunodeficiency virus type 1 envelope glycoprotein. J Virol. 1994 Mar;68(3):1689–1696. doi: 10.1128/jvi.68.3.1689-1696.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ehrlich L. S., Agresta B. E., Gelfand C. A., Jentoft J., Carter C. A. Spectral analysis and tryptic susceptibility as probes of HIV-1 capsid protein structure. Virology. 1994 Nov 1;204(2):515–525. doi: 10.1006/viro.1994.1565. [DOI] [PubMed] [Google Scholar]
- Franke E. K., Yuan H. E., Bossolt K. L., Goff S. P., Luban J. Specificity and sequence requirements for interactions between various retroviral Gag proteins. J Virol. 1994 Aug;68(8):5300–5305. doi: 10.1128/jvi.68.8.5300-5305.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Gelderblom H. R. Assembly and morphology of HIV: potential effect of structure on viral function. AIDS. 1991 Jun;5(6):617–637. [PubMed] [Google Scholar]
- 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]
- Guan K. L., Dixon J. E. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. doi: 10.1016/0003-2697(91)90534-z. [DOI] [PubMed] [Google Scholar]
- 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]
- Göttlinger H. G., Sodroski J. G., Haseltine W. A. Role of capsid precursor processing and myristoylation in morphogenesis and infectivity of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5781–5785. doi: 10.1073/pnas.86.15.5781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henderson L. E., Bowers M. A., Sowder R. C., 2nd, Serabyn S. A., Johnson D. G., Bess J. W., Jr, Arthur L. O., Bryant D. K., Fenselau C. Gag proteins of the highly replicative MN strain of human immunodeficiency virus type 1: posttranslational modifications, proteolytic processings, and complete amino acid sequences. J Virol. 1992 Apr;66(4):1856–1865. doi: 10.1128/jvi.66.4.1856-1865.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hughes B. P., Booth T. F., Belyaev A. S., McIlroy D., Jowett J., Roy P. Morphogenic capabilities of human immunodeficiency virus type 1 gag and gag-pol proteins in insect cells. Virology. 1993 Mar;193(1):242–255. doi: 10.1006/viro.1993.1120. [DOI] [PubMed] [Google Scholar]
- Jacks T., Power M. D., Masiarz F. R., Luciw P. A., Barr P. J., Varmus H. E. Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. Nature. 1988 Jan 21;331(6153):280–283. doi: 10.1038/331280a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Karayan L., Gay B., Gerfaux J., Boulanger P. A. Oligomerization of recombinant penton base of adenovirus type 2 and its assembly with fiber in baculovirus-infected cells. Virology. 1994 Aug 1;202(2):782–795. doi: 10.1006/viro.1994.1400. [DOI] [PubMed] [Google Scholar]
- 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]
- Langedijk J. P., Schalken J. J., Tersmette M., Huisman J. G., Meloen R. H. Location of epitopes on the major core protein p24 of human immunodeficiency virus. J Gen Virol. 1990 Nov;71(Pt 11):2609–2614. doi: 10.1099/0022-1317-71-11-2609. [DOI] [PubMed] [Google Scholar]
- Layne S. P., Merges M. J., Dembo M., Spouge J. L., Conley S. R., Moore J. P., Raina J. L., Renz H., Gelderblom H. R., Nara P. L. Factors underlying spontaneous inactivation and susceptibility to neutralization of human immunodeficiency virus. Virology. 1992 Aug;189(2):695–714. doi: 10.1016/0042-6822(92)90593-e. [DOI] [PubMed] [Google Scholar]
- Luban J., Bossolt K. L., Franke E. K., Kalpana G. V., Goff S. P. Human immunodeficiency virus type 1 Gag protein binds to cyclophilins A and B. Cell. 1993 Jun 18;73(6):1067–1078. doi: 10.1016/0092-8674(93)90637-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Luckow V. A., Summers M. D. High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology. 1989 May;170(1):31–39. doi: 10.1016/0042-6822(89)90348-6. [DOI] [PubMed] [Google Scholar]
- Löchelt M., Zentgraf H., Flügel R. M. Construction of an infectious DNA clone of the full-length human spumaretrovirus genome and mutagenesis of the bel 1 gene. Virology. 1991 Sep;184(1):43–54. doi: 10.1016/0042-6822(91)90820-2. [DOI] [PubMed] [Google Scholar]
- Mammano F., Ohagen A., Höglund S., Göttlinger H. G. Role of the major homology region of human immunodeficiency virus type 1 in virion morphogenesis. J Virol. 1994 Aug;68(8):4927–4936. doi: 10.1128/jvi.68.8.4927-4936.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mervis R. J., Ahmad N., Lillehoj E. P., Raum M. G., Salazar F. H., Chan H. W., Venkatesan S. The gag gene products of human immunodeficiency virus type 1: alignment within the gag open reading frame, identification of posttranslational modifications, and evidence for alternative gag precursors. J Virol. 1988 Nov;62(11):3993–4002. doi: 10.1128/jvi.62.11.3993-4002.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Novelli A., Boulanger P. A. Deletion analysis of functional domains in baculovirus-expressed adenovirus type 2 fiber. Virology. 1991 Nov;185(1):365–376. doi: 10.1016/0042-6822(91)90784-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Pal R., Reitz M. S., Jr, Tschachler E., Gallo R. C., Sarngadharan M. G., Veronese F. D. Myristoylation of gag proteins of HIV-1 plays an important role in virus assembly. AIDS Res Hum Retroviruses. 1990 Jun;6(6):721–730. doi: 10.1089/aid.1990.6.721. [DOI] [PubMed] [Google Scholar]
- Park J., Morrow C. D. The nonmyristylated Pr160gag-pol polyprotein of human immunodeficiency virus type 1 interacts with Pr55gag and is incorporated into viruslike particles. J Virol. 1992 Nov;66(11):6304–6313. doi: 10.1128/jvi.66.11.6304-6313.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rein A., McClure M. R., Rice N. R., Luftig R. B., Schultz A. M. Myristylation site in Pr65gag is essential for virus particle formation by Moloney murine leukemia virus. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7246–7250. doi: 10.1073/pnas.83.19.7246. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robert-Hebmann V., Emiliani S., Jean F., Resnicoff M., Traincard F., Devaux C. Clonal analysis of murine B cell response to the human immunodeficiency virus type 1 (HIV1)-gag p17 and p25 antigens. Mol Immunol. 1992 Jun;29(6):729–738. doi: 10.1016/0161-5890(92)90183-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Royer M., Hong S. S., Gay B., Cerutti M., Boulanger P. Expression and extracellular release of human immunodeficiency virus type 1 Gag precursors by recombinant baculovirus-infected cells. J Virol. 1992 May;66(5):3230–3235. doi: 10.1128/jvi.66.5.3230-3235.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schultz A. M., Rein A. Unmyristylated Moloney murine leukemia virus Pr65gag is excluded from virus assembly and maturation events. J Virol. 1989 May;63(5):2370–2373. doi: 10.1128/jvi.63.5.2370-2373.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith A. J., Srinivasakumar N., Hammarskjöld M. L., Rekosh D. Requirements for incorporation of Pr160gag-pol from human immunodeficiency virus type 1 into virus-like particles. J Virol. 1993 Apr;67(4):2266–2275. doi: 10.1128/jvi.67.4.2266-2275.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spearman P., Wang J. J., Vander Heyden N., Ratner L. Identification of human immunodeficiency virus type 1 Gag protein domains essential to membrane binding and particle assembly. J Virol. 1994 May;68(5):3232–3242. doi: 10.1128/jvi.68.5.3232-3242.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strambio-de-Castillia C., Hunter E. Mutational analysis of the major homology region of Mason-Pfizer monkey virus by use of saturation mutagenesis. J Virol. 1992 Dec;66(12):7021–7032. doi: 10.1128/jvi.66.12.7021-7032.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanchou V., Delaunay T., De Rocquigny H., Bodeus M., Darlix J. L., Roques B., Benarous R. Monoclonal antibody-mediated inhibition of RNA binding and annealing activities of HIV type 1 nucleocapsid protein. AIDS Res Hum Retroviruses. 1994 Aug;10(8):983–993. doi: 10.1089/aid.1994.10.983. [DOI] [PubMed] [Google Scholar]
- 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]
- Veronese F. D., Rahman R., Copeland T. D., Oroszlan S., Gallo R. C., Sarngadharan M. G. Immunological and chemical analysis of P6, the carboxyl-terminal fragment of HIV P15. AIDS Res Hum Retroviruses. 1987 Fall;3(3):253–264. doi: 10.1089/aid.1987.3.253. [DOI] [PubMed] [Google Scholar]
- Wagner R., Deml L., Fliessbach H., Wanner G., Wolf H. Assembly and extracellular release of chimeric HIV-1 Pr55gag retrovirus-like particles. Virology. 1994 Apr;200(1):162–175. doi: 10.1006/viro.1994.1175. [DOI] [PubMed] [Google Scholar]
- Wain-Hobson S., Sonigo P., Danos O., Cole S., Alizon M. Nucleotide sequence of the AIDS virus, LAV. Cell. 1985 Jan;40(1):9–17. doi: 10.1016/0092-8674(85)90303-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Wang C. T., Stegeman-Olsen J., Zhang Y., Barklis E. Assembly of HIV GAG-B-galactosidase fusion proteins into virus particles. Virology. 1994 May 1;200(2):524–534. doi: 10.1006/viro.1994.1215. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- Wills J. W. Retro-secretion of recombinant proteins. Nature. 1989 Jul 27;340(6231):323–324. doi: 10.1038/340323a0. [DOI] [PubMed] [Google Scholar]
- Yu X., Yu Q. C., Lee T. H., Essex M. The C terminus of human immunodeficiency virus type 1 matrix protein is involved in early steps of the virus life cycle. J Virol. 1992 Sep;66(9):5667–5670. doi: 10.1128/jvi.66.9.5667-5670.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Zhou W., Parent L. J., Wills J. W., Resh M. D. Identification of a membrane-binding domain within the amino-terminal region of human immunodeficiency virus type 1 Gag protein which interacts with acidic phospholipids. J Virol. 1994 Apr;68(4):2556–2569. doi: 10.1128/jvi.68.4.2556-2569.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]