Protein Science
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Protein Science (2005), 14:375-386. Published by Cold Spring Harbor Laboratory Press. Copyright © 2005 The Protein Society
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Morellet, N.
Right arrow Articles by Roques, B. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Morellet, N.
Right arrow Articles by Roques, B. P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Helical structure determined by NMR of the HIV-1 (345–392)Gag sequence, surrounding p2: Implications for particle assembly and RNA packaging

Nelly Morellet, Sabine Druillennec, Christine Lenoir, Serge Bouaziz and Bernard P. Roques

Unite de Pharmacologie Chimique et Genetique, Institut National de la Santé et la Recherche Medicale (INSERM) U640, Centre National de la Recherche Scientifique Unité Mixte de Recherche (CNRS UMR) 8151, Unité de Formation et de Recherche des Sciences Pharmaceutiques et Biologiques, 75270 Paris Cedex 06, France

(RECEIVED August 31, 2004; FINAL REVISION September 30, 2004; ACCEPTED September 30, 2004)

Gag protein oligomerization, an essential step during virus assembly, results in budding of spherical virus particles. This process is critically dependent on the spacer p2, located between the capsid and the nucleocapsid proteins. P2 contributes also, in association with NCp7, to specific recognition of the HIV-1 packaging signal resulting in viral genome encapsidation. There is no structural information about the 20 last amino acids of the C-terminal part of capsid (CA[CTD]) and p2, in the molecular mechanism of Gag assembly. In this study the structure of a peptide encompassing the 14 residues of p2 with the upstream 21 residues and the downstream 13 residues was determined by 1H NMR in 30% trifluoroethanol (TFE). The main structural motif is a well-defined amphipathic {alpha}-helix including p2, the seven last residues of the CA(CTD), and the two first residues of NCp7. Peptides containing the p2 domain have a strong tendency to aggregate in solution, as shown by gel filtration analyses in pure H2O. To take into account the aggregation phenomena, models of dimer and trimer formed through hydrophobic or hydrophilic interfaces were constructed by molecular dynamic simulations. Gel shift experiments demonstrate that the presence of at least p2 and the 13 first residues of NCp7 is required for RNA binding. A computer-generated model of the Gag polyprotein segment (282–434)Gag interacting with the packaging element SL3 is proposed, illustrating the importance of p2 and NCp7 in genomic encapsidation.

Keywords: Gag polyprotein; particle assembly; p2; HIV-1; NMR

Abbreviations: TFE, trifluoroethanol • HIV-1, human immunodeficiency virus type 1 • VLPs, virus-like particles • PR, protease • MA, matrix • CA, capsid • CA[CTD], C-terminal domain of CA • NCp7, nucleocapsid • DOSY, diffusion-ordered spectroscopy • cEM, cryoelectron microscopy • mGag, minimal Pr55Gag protein • RSV, Rous sarcoma virus • MLV, murine leukemia virus

Article and publication are at http://www.proteinscience.org/cgi/doi/10.1110/ps.041087605.

Reprint requests to: Nelly Morellet, Unite de Pharmacologie Chimique et Genetique, INSERM U640, CNRS UMR 8151, UFR des Sciences Pharmaceutiques et Biologiques, 4, Avenue de l’Observatoire, 75270 Paris Cedex 06, France; e-mail: morellet{at}pharmacie.univ-paris5.fr; fax: +(33)-1-43-26-69-18.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Virol.Home page
P. W. Keller, M. C. Johnson, and V. M. Vogt
Mutations in the Spacer Peptide and Adjoining Sequences in Rous Sarcoma Virus Gag Lead to Tubular Budding
J. Virol., July 15, 2008; 82(14): 6788 - 6797.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
C. S. Adamson, S. D. Ablan, I. Boeras, R. Goila-Gaur, F. Soheilian, K. Nagashima, F. Li, K. Salzwedel, M. Sakalian, C. T. Wild, et al.
In Vitro Resistance to the Human Immunodeficiency Virus Type 1 Maturation Inhibitor PA-457 (Bevirimat)
J. Virol., November 15, 2006; 80(22): 10957 - 10971.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
A. Ono, A. A. Waheed, A. Joshi, and E. O. Freed
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., November 15, 2005; 79(22): 14131 - 14140.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2005 by The Protein Society.