The tyrosine binding pocket in the adaptor protein 1 (AP-1) mu1 subunit is necessary for Nef to recruit AP-1 to the major histocompatibility complex class I cytoplasmic tail.
To evade the anti-human immunodeficiency virus (HIV) immune response, the HIV Nef protein disrupts major histocompatibility complex class I (MHC-I) trafficking by recruiting the clathrin adaptor protein 1 (AP-1) to the MHC-I cytoplasmic tail. Under normal conditions AP-1 binds dileucine and tyrosine signals (YXX phi motifs) via physically separate binding sites. In the case of the Nef-MHC-I complex, a tyrosine in the human leukocyte antigen (HLA)-A2 cytoplasmic tail ((320)YSQA) and a methionine in Nef (Met(20)) are absolutely required for AP-1 binding. Also present in Nef is a dileucine motif, which does not normally affect MHC-I trafficking and is not needed to recruit AP-1 to the Nef-MHC-I-complex. However, evidence is presented here that this dileucine motif can be activated by fusing Nef to the HLA-A2 tail in cis. Thus, the inability of this motif to function in trans likely results from a structural change that occurs when Nef binds to the MHC-I cytoplasmic tail. The physiologically relevant tyrosine-dependent recruitment of AP-1 to MHC-I, which occurs whether Nef is present in cis or trans, was stabilized by the acidic and polyproline domains within Nef. Additionally, amino acids Ala(324) and Asp(327) in the cytoplasmic tails of HLA-A and (but not HLA-C and HLA-E) molecules also stabilized AP-1 binding. Finally, mutation of the tyrosine binding pocket in the mu subunit of AP-1 created a dominant negative inhibitor of Nef-induced down-modulation of HLA-A2 that disrupted binding of wild type AP-1 to the Nef-MHC-I complex. Thus, these data provide evidence that Nef binding to the MHC-I cytoplasmic tail stabilizes the interaction of a tyrosine in the MHC-I cytoplasmic tail with the natural tyrosine binding pocket in AP-1.
Wonderlich
Elizabeth R
ER
Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, Michigan 48109.
Williams
Maya
M
Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, Michigan 48109.
Collins
Kathleen L
KL
Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, Michigan 48109; Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109; Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109. Electronic address: klcollin@umich.edu.
D033961:Adaptor Protein Complex 1 / Q000737:chemistry*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D003593:Cytoplasm / Q000378:metabolism; D005786:Gene Expression Regulation; D015693:Gene Products, nef / Q000737:chemistry*; D015395:Histocompatibility Antigens Class I / Q000737:chemistry*; D006801:Humans; D007930:Leucine / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014443:Tyrosine / Q000737:chemistry*
D033961:Adaptor Protein Complex 1; D015693:Gene Products, nef; D015395:Histocompatibility Antigens Class I; D014443:Tyrosine; D007930:Leucine