Nef binds p6* in GagPol during replication of HIV-1.
Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. TuOrA1138)
Costa LJ, Zheng YH, Mak J, Fackler OT, Peterlin BM UCSF, San Francisco, United States
The atypical Nef protein from HIV-1F12 (NefF12) 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 ER retention signal (NefKKXX). This mutant NefKKXX protein recapitulated fully the effects of NefF12 on Gag processing and virion production, either alone or as aCD8 fusion protein. Importantly, the mutant NefKKXX protein also localized to theIntermediate Compartment, between the Endoplasmic Reticulum and the TransGolgi 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 IntermediateCompartment interferes with viral replication and suggest a new role for Nef in the production of HIV-1.
Keywords: AEGIS, Gene Products, nef, Virus Replication, HIV-1, Virion, S100 Proteins, Genes, gag, S100A12 protein, human, In Vitro, Animal, metabolism, virology, genetics, pharmacokinetics
