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 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 arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lu, M.
Right arrow Articles by Nunberg, J. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lu, M.
Right arrow Articles by Nunberg, J. H.

 Previous Article  |  Next Article 

Journal of Virology, November 2001, p. 11146-11156, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11146-11156.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Structural and Functional Analysis of Interhelical Interactions in the Human Immunodeficiency Virus Type 1 gp41 Envelope Glycoprotein by Alanine-Scanning Mutagenesis

Min Lu,1,* Marisa O. Stoller,2 Shilong Wang,1 Jie Liu,1 Melinda B. Fagan,2,dagger and Jack H. Nunberg2,*

Department of Biochemistry, Weill Medical College of Cornell University, New York, New York 10021,1 and Montana Biotechnology Center, The University of Montana, Missoula, Montana 598122

Received 23 May 2001/Accepted 8 August 2001

Membrane fusion by human immunodeficiency virus type 1 (HIV-1) is promoted by the refolding of the viral envelope glycoprotein into a fusion-active conformation. The structure of the gp41 ectodomain core in its fusion-active state is a trimer of hairpins in which three antiparallel carboxyl-terminal helices pack into hydrophobic grooves on the surface of an amino-terminal trimeric coiled coil. In an effort to identify amino acid residues in these grooves that are critical for gp41 activation, we have used alanine-scanning mutagenesis to investigate the importance of individual side chains in determining the biophysical properties of the gp41 core and the membrane fusion activity of the gp120-gp41 complex. Alanine substitutions at Leu-556, Leu-565, Val-570, Gly-572, and Arg-579 positions severely impaired membrane fusion activity in envelope glycoproteins that were for the most part normally expressed. Whereas alanine mutations at Leu-565 and Val-570 destabilized the trimer-of-hairpins structure, mutations at Gly-572 and Arg-579 led to the formation of a stable gp41 core. Our results suggest that the Leu-565 and Val-570 residues are important determinants of conserved packing interactions between the amino- and carboxyl-terminal helices of gp41. We propose that the high degree of sequence conservation at Gly-572 and Arg-579 may result from selective pressures imposed by prefusogenic conformations of the HIV-1 envelope glycoprotein. Further analysis of the gp41 activation process may elucidate targets for antiviral intervention.

* Corresponding author. Mailing address for Jack H. Nunberg: Montana Biotechnology Center, The University of Montana, Missoula, MT 59812. Phone: (406) 243-6421. Fax: (406) 243-6425. E-mail: nunberg{at}selway.umt.edu. Mailing address for Min Lu: Department of Biochemistry, Weill Medical College of Cornell University, New York, NY 10021. Phone: (212) 746-6562. Fax: (212) 746-8875. E-mail: mlu{at}med.cornell.edu.

dagger Present address: Department of Philosophy, University of Texas, Austin, TX 78712.

Journal of Virology, November 2001, p. 11146-11156, Vol. 75, No. 22
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.22.11146-11156.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Markosyan, R. M., Leung, M. Y., Cohen, F. S. (2009). The Six-Helix Bundle of Human Immunodeficiency Virus Env Controls Pore Formation and Enlargement and Is Initiated at Residues Proximal to the Hairpin Turn. J. Virol. 83: 10048-10057 [Abstract] [Full Text]  
  • Eggink, D., Langedijk, J. P. M., Bonvin, A. M. J. J., Deng, Y., Lu, M., Berkhout, B., Sanders, R. W. (2009). Detailed Mechanistic Insights into HIV-1 Sensitivity to Three Generations of Fusion Inhibitors. J. Biol. Chem. 284: 26941-26950 [Abstract] [Full Text]  
  • He, Y., Liu, S., Li, J., Lu, H., Qi, Z., Liu, Z., Debnath, A. K., Jiang, S. (2008). Conserved Salt Bridge between the N- and C-Terminal Heptad Repeat Regions of the Human Immunodeficiency Virus Type 1 gp41 Core Structure Is Critical for Virus Entry and Inhibition. J. Virol. 82: 11129-11139 [Abstract] [Full Text]  
  • He, Y., Cheng, J., Lu, H., Li, J., Hu, J., Qi, Z., Liu, Z., Jiang, S., Dai, Q. (2008). Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains. Proc. Natl. Acad. Sci. USA 105: 16332-16337 [Abstract] [Full Text]  
  • He, Y., Cheng, J., Li, J., Qi, Z., Lu, H., Dong, M., Jiang, S., Dai, Q. (2008). Identification of a Critical Motif for the Human Immunodeficiency Virus Type 1 (HIV-1) gp41 Core Structure: Implications for Designing Novel Anti-HIV Fusion Inhibitors. J. Virol. 82: 6349-6358 [Abstract] [Full Text]  
  • He, Y., Liu, S., Jing, W., Lu, H., Cai, D., Chin, D. J., Debnath, A. K., Kirchhoff, F., Jiang, S. (2007). Conserved Residue Lys574 in the Cavity of HIV-1 Gp41 Coiled-coil Domain Is Critical for Six-helix Bundle Stability and Virus Entry. J. Biol. Chem. 282: 25631-25639 [Abstract] [Full Text]  
  • Chinnadurai, R., Rajan, D., Munch, J., Kirchhoff, F. (2007). Human Immunodeficiency Virus Type 1 Variants Resistant to First- and Second-Version Fusion Inhibitors and Cytopathic in Ex Vivo Human Lymphoid Tissue. J. Virol. 81: 6563-6572 [Abstract] [Full Text]  
  • Doyle, J., Prussia, A., White, L. K., Sun, A., Liotta, D. C., Snyder, J. P., Compans, R. W., Plemper, R. K. (2006). Two Domains That Control Prefusion Stability and Transport Competence of the Measles Virus Fusion Protein. J. Virol. 80: 1524-1536 [Abstract] [Full Text]  
  • Suntoke, T. R., Chan, D. C. (2005). The Fusion Activity of HIV-1 gp41 Depends on Interhelical Interactions. J. Biol. Chem. 280: 19852-19857 [Abstract] [Full Text]  
  • Kinomoto, M., Yokoyama, M., Sato, H., Kojima, A., Kurata, T., Ikuta, K., Sata, T., Tokunaga, K. (2005). Amino Acid 36 in the Human Immunodeficiency Virus Type 1 gp41 Ectodomain Controls Fusogenic Activity: Implications for the Molecular Mechanism of Viral Escape from a Fusion Inhibitor. J. Virol. 79: 5996-6004 [Abstract] [Full Text]  
  • Heap, C. J., Reading, S. A., Dimmock, N. J. (2005). An antibody specific for the C-terminal tail of the gp41 transmembrane protein of human immunodeficiency virus type 1 mediates post-attachment neutralization, probably through inhibition of virus-cell fusion. J. Gen. Virol. 86: 1499-1507 [Abstract] [Full Text]  
  • Desmezieres, E., Gupta, N., Vassell, R., He, Y., Peden, K., Sirota, L., Yang, Z., Wingfield, P., Weiss, C. D. (2005). Human Immunodeficiency Virus (HIV) gp41 Escape Mutants: Cross-Resistance to Peptide Inhibitors of HIV Fusion and Altered Receptor Activation of gp120. J. Virol. 79: 4774-4781 [Abstract] [Full Text]  
  • West, D. S., Sheehan, M. S., Segeleon, P. K., Dutch, R. E. (2005). Role of the Simian Virus 5 Fusion Protein N-Terminal Coiled-Coil Domain in Folding and Promotion of Membrane Fusion. J. Virol. 79: 1543-1551 [Abstract] [Full Text]  
  • York, J., Nunberg, J. H. (2004). Role of Hydrophobic Residues in the Central Ectodomain of gp41 in Maintaining the Association between Human Immunodeficiency Virus Type 1 Envelope Glycoprotein Subunits gp120 and gp41. J. Virol. 78: 4921-4926 [Abstract] [Full Text]  
  • Murakami, T., Ablan, S., Freed, E. O., Tanaka, Y. (2004). Regulation of Human Immunodeficiency Virus Type 1 Env-Mediated Membrane Fusion by Viral Protease Activity. J. Virol. 78: 1026-1031 [Abstract] [Full Text]  
  • Russell, C. J., Kantor, K. L., Jardetzky, T. S., Lamb, R. A. (2003). A dual-functional paramyxovirus F protein regulatory switch segment: activation and membrane fusion. JCB 163: 363-374 [Abstract] [Full Text]  
  • Sia, S. K., Kim, P. S. (2003). Protein grafting of an HIV-1-inhibiting epitope. Proc. Natl. Acad. Sci. USA 100: 9756-9761 [Abstract] [Full Text]  
  • Giannecchini, S., Di Fenza, A., D'Ursi, A. M., Matteucci, D., Rovero, P., Bendinelli, M. (2003). Antiviral Activity and Conformational Features of an Octapeptide Derived from the Membrane-Proximal Ectodomain of the Feline Immunodeficiency Virus Transmembrane Glycoprotein. J. Virol. 77: 3724-3733 [Abstract] [Full Text]  
  • Markosyan, R. M., Cohen, F. S., Melikyan, G. B. (2003). HIV-1 Envelope Proteins Complete Their Folding into Six-helix Bundles Immediately after Fusion Pore Formation. Mol. Biol. Cell 14: 926-938 [Abstract] [Full Text]  
  • Sanders, R. W., Vesanen, M., Schuelke, N., Master, A., Schiffner, L., Kalyanaraman, R., Paluch, M., Berkhout, B., Maddon, P. J., Olson, W. C., Lu, M., Moore, J. P. (2002). Stabilization of the Soluble, Cleaved, Trimeric Form of the Envelope Glycoprotein Complex of Human Immunodeficiency Virus Type 1. J. Virol. 76: 8875-8889 [Abstract] [Full Text]  
  • Follis, K. E., Larson, S. J., Lu, M., Nunberg, J. H. (2002). Genetic Evidence that Interhelical Packing Interactions in the gp41 Core Are Critical for Transition of the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein to the Fusion-Active State. J. Virol. 76: 7356-7362 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»