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Journal of Virology, May 2004, p. 5311-5323, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5311-5323.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Nef Binds p6* in GagPol during Replication of Human Immunodeficiency Virus Type 1

Luciana J. Costa,1 Yong-Hui Zheng,1 Jerica Sabotic,1 Johnson Mak,2 Oliver T. Fackler,3* and B. Matija Peterlin1*

Departments of Medicine, Microbiology, and Immunology, University of California, San Francisco, California 94143-0703,1 AIDS Pathogenesis Research Unit, The Macfarlane Burnet Institute for Medical Research and Public Health, and Department of Biochemistry and Molecular Biology, Monash University, Clayton, Australia,2 Abteilung Virologie, Universitätsklinikum Heidelberg, Heidelberg, Germany3

Received 9 November 2003/ Accepted 12 January 2004

The atypical Nef protein (NefF12) from human immunodeficiency virus type 1 strain F12 (HIV-1F12) interferes with virion production and infectivity via a mysterious mechanism. The correlation of these effects with the unusual perinuclear subcellular localization of NefF12 suggested that the wild-type Nef protein could bind to assembly intermediates in late stages of viral replication. To test this hypothesis, Nef from HIV-1NL4-3 was fused to an endoplasmic reticulum (ER) retention signal (NefKKXX). This mutant NefKKXX protein recapitulated fully the effects of NefF12 on Gag processing and virion production, either alone or as a CD8 fusion protein. Importantly, the mutant NefKKXX protein also localized to the intermediate compartment, between the ER and the trans-Golgi network. Furthermore, Nef bound the GagPol polyprotein in vitro and in vivo. This binding mapped to the C-terminal flexible loop in Nef and the transframe p6* protein in GagPol. The significance of this interaction was demonstrated by a genetic assay in which the release of a mutant HIV-1 provirus lacking the PTAP motif in the late domain that no longer binds Tsg101 was rescued by a Nef.Tsg101 chimera. Importantly, this rescue as well as incorporation of Nef into HIV-1 virions correlated with the ability of Nef to interact with GagPol. Our data demonstrate that the retention of Nef in the intermediate compartment interferes with viral replication and suggest a new role for Nef in the production of HIV-1.

* Corresponding author. Mailing address for B. Matija Peterlin: Room N215, UCSF-Mt. Zion Cancer Center, 2340 Sutter St., San Francisco, CA 94115. Phone: (415) 502-1905. Fax: (415) 502-1901. E-mail: matija{at}itsa.ucsf.edu. Mailing address for Oliver T. Fackler: Abteilung Virologie, Universitätsklinikum Heidelberg, INF 324, D-69120 Heidelberg, Germany. Phone: 49-(0)6221-565007. Fax: 49-(0)6221-565003. E-mail: oliver_fackler{at}med.uni-heidelberg.de.

Journal of Virology, May 2004, p. 5311-5323, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5311-5323.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Leiherer, A., Ludwig, C., Wagner, R. (2009). Uncoupling Human Immunodeficiency Virus Type 1 gag and pol Reading Frames: Role of the Transframe Protein p6* in Viral Replication. J. Virol. 83: 7210-7220 [Abstract] [Full Text]  
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