Previous Article | Next Article 
Journal of Virology, November 2002, p. 11729-11737, Vol. 76, No. 22
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.22.11729-11737.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Characterization of a Putative
-Helix across the Capsid-SP1 Boundary That Is Critical for the Multimerization of Human Immunodeficiency Virus Type 1 Gag
Chen Liang,1,2* Jing Hu,1 Rodney S. Russell,1,3 Ariel Roldan,1 Lawrence Kleiman,1,2,3 and Mark A. Wainberg1,2,3
McGill AIDS Centre, Lady Davis Institute-Jewish General Hospital, Montreal, Quebec, Canada H3T 1E2,1
Departments of Medicine,2
Microbiology & Immunology, McGill University, Montreal, Quebec, Canada H3A 2B43
Received 17 May 2002/
Accepted 7 August 2002
A 14-amino-acid spacer peptide termed SP1 that separates the capsid (CA) and nucleocapsid (NC) sequences plays an active role in the assembly of human immunodeficiency virus type 1. This activity of SP1 involves its amino-terminal residues that, together with adjacent CA residues, constitute a putative
-helical structure spanning Gag residues from positions 359 to 371. In this study, we have determined that the virus assembly determinants within this putative
-helix were residues H359, K360, A361, L364, A367, and M368, of which K360 and A367 contribute to virus production to lesser extents. Notably, changes of the two basic amino acids H359 and K360 to arginine (R) impaired virus production, whereas mutations L364I and M368I, in contrast to L364A and M368A, generated near-wild-type levels of virus particles. This suggests that within Gag complexes, amino acids H359 and K360 are involved in stricter steric interactions than L364 and M368. Since L364 and M368 are separated by four residues and thus presumably located on the same side of the helical surface, they may initiate synergistic hydrophobic interactions to stabilize Gag association. Further analysis in the context of the protease-negative mutation D185H confirmed the key roles of amino acids H359, A361, L364, and M368 in virus assembly. Importantly, when transfected cells were subjected to Dounce homogenization and the cell lysates were treated by ultracentrifugation at 100,000 x g, Gag molecules containing each of the H359A, A361V, L364A, and M368A mutations were found mainly in the supernatant fraction (S100), whereas approximately 80% of wild-type Gag proteins were found in the pellet. Therefore, these four mutations must have prevented Gag from generating large complexes.
* Corresponding author. Mailing address: McGill AIDS Centre, Lady Davis Institute-Jewish General Hospital, 3755 Cote Ste-Catherine Rd., Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7537. E-mail:
chen.liang{at}mcgill.ca.
Journal of Virology, November 2002, p. 11729-11737, Vol. 76, No. 22
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.22.11729-11737.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
This article has been cited by other articles:
-
Briggs, J. A. G., Riches, J. D., Glass, B., Bartonova, V., Zanetti, G., Krausslich, H.-G.
(2009). Structure and assembly of immature HIV. Proc. Natl. Acad. Sci. USA
106: 11090-11095
[Abstract]
[Full Text]
-
Heslin, D. J., Murcia, P., Arnaud, F., Van Doorslaer, K., Palmarini, M., Lenz, J.
(2009). A Single Amino Acid Substitution in a Segment of the CA Protein within Gag That Has Similarity to Human Immunodeficiency Virus Type 1 Blocks Infectivity of a Human Endogenous Retrovirus K Provirus in the Human Genome. J. Virol.
83: 1105-1114
[Abstract]
[Full Text]
-
Keller, P. W., Johnson, M. C., Vogt, V. M.
(2008). Mutations in the Spacer Peptide and Adjoining Sequences in Rous Sarcoma Virus Gag Lead to Tubular Budding. J. Virol.
82: 6788-6797
[Abstract]
[Full Text]
-
Chatel-Chaix, L., Abrahamyan, L., Frechina, C., Mouland, A. J., DesGroseillers, L.
(2007). The Host Protein Staufen1 Participates in Human Immunodeficiency Virus Type 1 Assembly in Live Cells by Influencing pr55Gag Multimerization. J. Virol.
81: 6216-6230
[Abstract]
[Full Text]
-
Liao, W.-H., Huang, K.-J., Chang, Y.-F., Wang, S.-M., Tseng, Y.-T., Chiang, C.-C., Wang, J.-J., Wang, C.-T.
(2007). Incorporation of Human Immunodeficiency Virus Type 1 Reverse Transcriptase into Virus-Like Particles. J. Virol.
81: 5155-5165
[Abstract]
[Full Text]
-
Zhou, J., Chen, C. H., Aiken, C.
(2006). Human Immunodeficiency Virus Type 1 Resistance to the Small Molecule Maturation Inhibitor 3-O-(3',3'-Dimethylsuccinyl)-Betulinic Acid Is Conferred by a Variety of Single Amino Acid Substitutions at the CA-SP1 Cleavage Site in Gag. J. Virol.
80: 12095-12101
[Abstract]
[Full Text]
-
Adamson, C. S., Ablan, S. D., Boeras, I., Goila-Gaur, R., Soheilian, F., Nagashima, K., Li, F., Salzwedel, K., Sakalian, M., Wild, C. T., Freed, E. O.
(2006). In Vitro Resistance to the Human Immunodeficiency Virus Type 1 Maturation Inhibitor PA-457 (Bevirimat). J. Virol.
80: 10957-10971
[Abstract]
[Full Text]
-
Saad, J. S., Miller, J., Tai, J., Kim, A., Ghanam, R. H., Summers, M. F.
(2006). From the Cover: Structural basis for targeting HIV-1 Gag proteins to the plasma membrane for virus assembly. Proc. Natl. Acad. Sci. USA
103: 11364-11369
[Abstract]
[Full Text]
-
Ono, A., Waheed, A. A., Joshi, A., Freed, E. O.
(2005). Association of Human Immunodeficiency Virus Type 1 Gag with Membrane Does Not Require Highly Basic Sequences in the Nucleocapsid: Use of a Novel Gag Multimerization Assay. J. Virol.
79: 14131-14140
[Abstract]
[Full Text]
-
Ako-Adjei, D., Johnson, M. C., Vogt, V. M.
(2005). The Retroviral Capsid Domain Dictates Virion Size, Morphology, and Coassembly of Gag into Virus-Like Particles. J. Virol.
79: 13463-13472
[Abstract]
[Full Text]
-
Roldan, A., Warren, O. U., Russell, R. S., Liang, C., Wainberg, M. A.
(2005). A HIV-1 Minimal Gag Protein Is Superior to Nucleocapsid at in Vitro Annealing and Exhibits Multimerization-induced Inhibition of Reverse Transcription. J. Biol. Chem.
280: 17488-17496
[Abstract]
[Full Text]
-
Guo, X., Roldan, A., Hu, J., Wainberg, M. A., Liang, C.
(2005). Mutation of the SP1 Sequence Impairs both Multimerization and Membrane-Binding Activities of Human Immunodeficiency Virus Type 1 Gag. J. Virol.
79: 1803-1812
[Abstract]
[Full Text]
-
Marozsan, A. J., Fraundorf, E., Abraha, A., Baird, H., Moore, D., Troyer, R., Nankja, I., Arts, E. J.
(2004). Relationships between Infectious Titer, Capsid Protein Levels, and Reverse Transcriptase Activities of Diverse Human Immunodeficiency Virus Type 1 Isolates. J. Virol.
78: 11130-11141
[Abstract]
[Full Text]
-
Roldan, A., Russell, R. S., Marchand, B., Gotte, M., Liang, C., Wainberg, M. A.
(2004). In Vitro Identification and Characterization of an Early Complex Linking HIV-1 Genomic RNA Recognition and Pr55Gag Multimerization. J. Biol. Chem.
279: 39886-39894
[Abstract]
[Full Text]
-
Melamed, D., Mark-Danieli, M., Kenan-Eichler, M., Kraus, O., Castiel, A., Laham, N., Pupko, T., Glaser, F., Ben-Tal, N., Bacharach, E.
(2004). The Conserved Carboxy Terminus of the Capsid Domain of Human Immunodeficiency Virus Type 1 Gag Protein Is Important for Virion Assembly and Release. J. Virol.
78: 9675-9688
[Abstract]
[Full Text]
-
Guo, X., Hu, J., Whitney, J. B., Russell, R. S., Liang, C.
(2004). Important Role for the CA-NC Spacer Region in the Assembly of Bovine Immunodeficiency Virus Gag Protein. J. Virol.
78: 551-560
[Abstract]
[Full Text]
-
Tang, C., Loeliger, E., Luncsford, P., Kinde, I., Beckett, D., Summers, M. F.
(2004). From the Cover: Entropic switch regulates myristate exposure in the HIV-1 matrix protein. Proc. Natl. Acad. Sci. USA
101: 517-522
[Abstract]
[Full Text]
-
Li, F., Goila-Gaur, R., Salzwedel, K., Kilgore, N. R., Reddick, M., Matallana, C., Castillo, A., Zoumplis, D., Martin, D. E., Orenstein, J. M., Allaway, G. P., Freed, E. O., Wild, C. T.
(2003). PA-457: A potent HIV inhibitor that disrupts core condensation by targeting a late step in Gag processing. Proc. Natl. Acad. Sci. USA
100: 13555-13560
[Abstract]
[Full Text]
-
von Schwedler, U. K., Stray, K. M., Garrus, J. E., Sundquist, W. I.
(2003). Functional Surfaces of the Human Immunodeficiency Virus Type 1 Capsid Protein. J. Virol.
77: 5439-5450
[Abstract]
[Full Text]