Two Independent Regions of HIV‐1 Nef are Required for Connection with the Endocytic Pathway Through Binding to the μ1 Chain of AP1 Complex

The Nef protein from the human immunodeficiency virus (HIV) induces down‐regulation of the CD4 and major histocompatibility complex class I molecules from the cell surface by interfering with the endocytic machinery. This work focuses on the interaction of HIV‐1 Nef with the μ1 chain of adaptor protein type 1 (AP1) complex and its contribution to the Nef‐induced alterations of membrane trafficking. Two independent regions surrounding a disordered loop located in the C‐terminal part of Nef are involved in μ1 binding. Each region can separately interact with μ1, and simultaneous point mutations within both regions are needed to abolish binding. We used CD8 chimeras in which the cytoplasmic tail was replaced by Nef mutants to show that these μ1‐binding sites contain determinants required to induce CD4 down‐regulation and to target the chimera to the endocytic pathway by promoting AP1 complex recruitment. Ultrastructural analysis revealed that the CD8‐Nef chimera provokes morphological alterations of the endosomal compartments and co‐localizes with AP1 complexes. These data indicate that the recruitment by Nef of AP1 via binding to μ1 participates in the connection of Nef with the endocytic pathway.

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