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 Welker, R.
Right arrow Articles by Kräusslich, H.-G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Welker, R.
Right arrow Articles by Kräusslich, H.-G.

 Previous Article  |  Next Article 

Journal of Virology, November 1998, p. 8833-8840, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Virion Incorporation of Human Immunodeficiency Virus Type 1 Nef Is Mediated by a Bipartite Membrane-Targeting Signal: Analysis of Its Role in Enhancement of Viral Infectivity

Reinhold Welker,1 Mark Harris,2 Bettina Cardel,1 and Hans-Georg Kräusslich1,*

Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, D-20251 Hamburg, Germany,1 and Department of Microbiology, University of Leeds, Leeds LS2 9JT, United Kingdom2

Received 24 March 1998/Accepted 27 July 1998

The nef gene of primate immunodeficiency viruses is essential for high-titer virus replication and AIDS pathogenesis in vivo. In tissue culture, Nef is not required for human immunodeficiency virus (HIV) infection but enhances viral infectivity. We and others have shown that Nef is incorporated into HIV-1 particles and cleaved by the viral proteinase. To determine the signal for Nef incorporation and to analyze whether virion-associated Nef is responsible for enhancement of infectivity, we generated a panel of nef mutants and analyzed them for virion incorporation of Nef and for their relative infectivities. We report that N-terminal truncations of Nef abolished its incorporation into HIV particles. Incorporation was reconstituted by targeting the respective proteins to the plasma membrane by using a heterologous signal. Mutational analysis revealed that both myristoylation and an N-terminal cluster of basic amino acids were required for virion incorporation and for plasma membrane targeting of Nef. Grafting the N-terminal anchor domain of Nef onto the green fluorescent protein led to membrane targeting and virion incorporation of the resulting fusion protein. These results indicate that Nef incorporation into HIV-1 particles is mediated by plasma membrane targeting via an N-terminal bipartite signal which is reminiscent of a Src homology region 4. Virion incorporation of Nef correlated with enhanced infectivity of the respective viruses in a single-round replication assay. However, the phenotypes of HIV mutants with reduced Nef incorporation only partly correlated with their ability to replicate in primary lymphocytes, indicating that additional or different mechanisms may be involved in this system.

* Corresponding author. Mailing address: Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, Martinistr. 52, D-20251 Hamburg, Germany. Phone: 49 40 48051-241. Fax: 49 40 48051-184. E-mail: hgk{at}hpi.uni-hamburg.de.

Journal of Virology, November 1998, p. 8833-8840, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Van Rhijn, I., Young, D. C., De Jong, A., Vazquez, J., Cheng, T.-Y., Talekar, R., Barral, D. C., Leon, L., Brenner, M. B., Katz, J. T., Riese, R., Ruprecht, R. M., O'Connor, P. B., Costello, C. E., Porcelli, S. A., Briken, V., Moody, D. B. (2009). CD1c bypasses lysosomes to present a lipopeptide antigen with 12 amino acids. JEM 206: 1409-1422 [Abstract] [Full Text]  
  • Green, L. A., Liu, Y., He, J. J. (2009). Inhibition of HIV-1 Infection and Replication by Enhancing Viral Incorporation of Innate Anti-HIV-1 Protein A3G: A NON-PATHOGENIC Nef MUTANT-BASED ANTI-HIV STRATEGY. J. Biol. Chem. 284: 13363-13372 [Abstract] [Full Text]  
  • Laguette, N., Benichou, S., Basmaciogullari, S. (2009). Human Immunodeficiency Virus Type 1 Nef Incorporation into Virions Does Not Increase Infectivity. J. Virol. 83: 1093-1104 [Abstract] [Full Text]  
  • Seaton, K. E., Smith, C. D. (2008). N-Myristoyltransferase isozymes exhibit differential specificity for human immunodeficiency virus type 1 Gag and Nef. J. Gen. Virol. 89: 288-296 [Abstract] [Full Text]  
  • Pizzato, M., Helander, A., Popova, E., Calistri, A., Zamborlini, A., Palu, G., Gottlinger, H. G. (2007). Dynamin 2 is required for the enhancement of HIV-1 infectivity by Nef. Proc. Natl. Acad. Sci. USA 104: 6812-6817 [Abstract] [Full Text]  
  • Mangino, G., Percario, Z. A., Fiorucci, G., Vaccari, G., Manrique, S., Romeo, G., Federico, M., Geyer, M., Affabris, E. (2007). In Vitro Treatment of Human Monocytes/Macrophages with Myristoylated Recombinant Nef of Human Immunodeficiency Virus Type 1 Leads to the Activation of Mitogen-Activated Protein Kinases, I{kappa}B Kinases, and Interferon Regulatory Factor 3 and to the Release of Beta Interferon. J. Virol. 81: 2777-2791 [Abstract] [Full Text]  
  • Qi, M., Aiken, C. (2007). Selective Restriction of Nef-Defective Human Immunodeficiency Virus Type 1 by a Proteasome-Dependent Mechanism. J. Virol. 81: 1534-1536 [Abstract] [Full Text]  
  • Bentham, M., Mazaleyrat, S., Harris, M. (2006). Role of myristoylation and N-terminal basic residues in membrane association of the human immunodeficiency virus type 1 Nef protein.. J. Gen. Virol. 87: 563-571 [Abstract] [Full Text]  
  • Yu, Z., Sanchez-Velar, N., Catrina, I. E., Kittler, E. L. W., Udofia, E. B., Zapp, M. L. (2005). The cellular HIV-1 Rev cofactor hRIP is required for viral replication. Proc. Natl. Acad. Sci. USA 102: 4027-4032 [Abstract] [Full Text]  
  • Bohne, J., Wodrich, H., Kräusslich, H.-G. (2005). Splicing of human immunodeficiency virus RNA is position-dependent suggesting sequential removal of introns from the 5' end. Nucleic Acids Res 33: 825-837 [Abstract] [Full Text]  
  • Hill, B. T., Skowronski, J. (2005). Human N-Myristoyltransferases Form Stable Complexes with Lentiviral Nef and Other Viral and Cellular Substrate Proteins. J. Virol. 79: 1133-1141 [Abstract] [Full Text]  
  • Campbell, E. M., Nunez, R., Hope, T. J. (2004). Disruption of the Actin Cytoskeleton Can Complement the Ability of Nef To Enhance Human Immunodeficiency Virus Type 1 Infectivity. J. Virol. 78: 5745-5755 [Abstract] [Full Text]  
  • Alexander, M., Bor, Y.-c., Ravichandran, K. S., Hammarskjold, M.-L., Rekosh, D. (2004). Human Immunodeficiency Virus Type 1 Nef Associates with Lipid Rafts To Downmodulate Cell Surface CD4 and Class I Major Histocompatibility Complex Expression and To Increase Viral Infectivity. J. Virol. 78: 1685-1696 [Abstract] [Full Text]  
  • Chazal, N., Gerlier, D. (2003). Virus Entry, Assembly, Budding, and Membrane Rafts. Microbiol. Mol. Biol. Rev. 67: 226-237 [Abstract] [Full Text]  
  • Forshey, B. M., Aiken, C. (2003). Disassembly of Human Immunodeficiency Virus Type 1 Cores In Vitro Reveals Association of Nef with the Subviral Ribonucleoprotein Complex. J. Virol. 77: 4409-4414 [Abstract] [Full Text]  
  • Khan, M., Garcia-Barrio, M., Powell, M. D. (2003). Treatment of Human Immunodeficiency Virus Type 1 Virions Depleted of Cyclophilin A by Natural Endogenous Reverse Transcription Restores Infectivity. J. Virol. 77: 4431-4434 [Abstract] [Full Text]  
  • Chakrabarti, L. A., Metzner, K. J., Ivanovic, T., Cheng, H., Louis-Virelizier, J., Connor, R. I., Cheng-Mayer, C. (2002). A Truncated Form of Nef Selected during Pathogenic Reversion of Simian Immunodeficiency Virus SIVmac239{Delta}nef Increases Viral Replication. J. Virol. 77: 1245-1256 [Abstract] [Full Text]  
  • Dorfman, T., Popova, E., Pizzato, M., Gottlinger, H. G. (2002). Nef Enhances Human Immunodeficiency Virus Type 1 Infectivity in the Absence of Matrix. J. Virol. 76: 6857-6862 [Abstract] [Full Text]  
  • Venkatesan, S., Petrovic, A., Locati, M., Kim, Y.-O., Weissman, D., Murphy, P. M. (2001). A Membrane-proximal Basic Domain and Cysteine Cluster in the C-terminal Tail of CCR5 Constitute a Bipartite Motif Critical for Cell Surface Expression. J. Biol. Chem. 276: 40133-40145 [Abstract] [Full Text]  
  • Zhou, J., Aiken, C. (2001). Nef Enhances Human Immunodeficiency Virus Type 1 Infectivity Resulting from Intervirion Fusion: Evidence Supporting a Role for Nef at the Virion Envelope. J. Virol. 75: 5851-5859 [Abstract] [Full Text]  
  • Schaeffer, E., Geleziunas, R., Greene, W. C. (2001). Human Immunodeficiency Virus Type 1 Nef Functions at the Level of Virus Entry by Enhancing Cytoplasmic Delivery of Virions. J. Virol. 75: 2993-3000 [Abstract] [Full Text]  
  • Manninen, A., Huotari, P., Hiipakka, M., Renkema, G. H., Saksela, K. (2001). Activation of NFAT-Dependent Gene Expression by Nef: Conservation among Divergent Nef Alleles, Dependence on SH3 Binding and Membrane Association, and Cooperation with Protein Kinase C-{theta}. J. Virol. 75: 3034-3037 [Abstract] [Full Text]  
  • Hildinger, M., Dittmar, M. T., Schult-Dietrich, P., Fehse, B., Schnierle, B. S., Thaler, S., Stiegler, G., Welker, R., von Laer, D. (2001). Membrane-Anchored Peptide Inhibits Human Immunodeficiency Virus Entry. J. Virol. 75: 3038-3042 [Abstract] [Full Text]  
  • Akari, H., Arold, S., Fukumori, T., Okazaki, T., Strebel, K., Adachi, A. (2000). Nef-Induced Major Histocompatibility Complex Class I Down-Regulation Is Functionally Dissociated from Its Virion Incorporation, Enhancement of Viral Infectivity, and CD4 Down-Regulation. J. Virol. 74: 2907-2912 [Abstract] [Full Text]  
  • Welker, R., Hohenberg, H., Tessmer, U., Huckhagel, C., Kräusslich, H.-G. (2000). Biochemical and Structural Analysis of Isolated Mature Cores of Human Immunodeficiency Virus Type 1. J. Virol. 74: 1168-1177 [Abstract] [Full Text]  
  • Kotov, A., Zhou, J., Flicker, P., Aiken, C. (1999). Association of Nef with the Human Immunodeficiency Virus Type 1 Core. J. Virol. 73: 8824-8830 [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»