Mechanism of Nef‐induced CD4 endocytosis: Nef connects CD4 with the μ chain of adaptor complexes

The Nef protein of primate lentiviruses down‐regulates the cell surface expression of CD4 and probably MHC I by connecting these receptors with the endocytic machinery. Here, we reveal that Nef interacts with the μ chains of adaptor complexes, key components of clathrin‐coated pits. For human immunodeficiency virus type 2 (HIV‐2) and simian immunodeficiency virus (SIV) Nef, this interaction occurs via tyrosine‐based motifs reminiscent of endocytosis signals. Mutating these motifs prevents the binding of SIV Nef to the μ chain of plasma membrane adaptor complexes, abrogates its ability to induce CD4 internalization, suppresses the accelerated endocytosis of a chimeric integral membrane protein harboring Nef as its cytoplasmic domain and confers a dominant‐negative phenotype to the viral protein. Taken together, these data identify μ adaptins as downstream mediators of the down‐modulation of CD4, and possibly MHC I, by Nef.

[1]  B. Walker,et al.  HIV-1 Nef protein protects infected primary cells against killing by cytotoxic T lymphocytes , 1998, Nature.

[2]  K. Krause,et al.  Nef-mediated Clathrin-coated Pit Formation , 1997, The Journal of cell biology.

[3]  Scott D Emr,et al.  The AP-3 Adaptor Complex Is Essential for Cargo-Selective Transport to the Yeast Vacuole , 1997, Cell.

[4]  C. Geisler,et al.  Regulation and Function of the CD3γ DxxxLL Motif: A Binding Site for Adaptor Protein-1 and Adaptor Protein-2 in Vitro , 1997, The Journal of cell biology.

[5]  L. Cantley,et al.  Regulatory interactions in the recognition of endocytic sorting signals by AP‐2 complexes , 1997, The EMBO journal.

[6]  J. Bonifacino,et al.  Tyrosine phosphorylation controls internalization of CTLA-4 by regulating its interaction with clathrin-associated adaptor complex AP-2. , 1997, Immunity.

[7]  A. Verkleij,et al.  Association and Colocalization of Eps15 with Adaptor Protein-2 and Clathrin , 1997, The Journal of cell biology.

[8]  J. Skowroński,et al.  Separable functions of Nef disrupt two aspects of T cell receptor machinery: CD4 expression and CD3 signaling , 1997, The EMBO journal.

[9]  M. Foti,et al.  The HIV-1 Nef protein acts as a connector with sorting pathways in the Golgi and at the plasma membrane. , 1997, Immunity.

[10]  J. Bonifacino,et al.  Structural Determinants of Interaction of Tyrosine-based Sorting Signals with the Adaptor Medium Chains* , 1996, The Journal of Biological Chemistry.

[11]  S. Grzesiek,et al.  The CD4 determinant for downregulation by HIV-1 Nef directly binds to Nef. Mapping of the Nef binding surface by NMR. , 1996, Biochemistry.

[12]  A. Dautry‐Varsat,et al.  The Ear of -Adaptin Interacts with the COOH-terminal Domain of the Eps15 Protein (*) , 1996, The Journal of Biological Chemistry.

[13]  D. Trono,et al.  Mutational analysis of HIV-1 Nef: identification of two mutants that are temperature-sensitive for CD4 downregulation. , 1996, Virology.

[14]  G. Milanesi,et al.  Nef-CD4 physical interaction sensed with the yeast two-hybrid system. , 1996, Virology.

[15]  Y. Kido,et al.  Interaction of Shc with Adaptor Protein Adaptins (*) , 1996, The Journal of Biological Chemistry.

[16]  F. Lemonnier,et al.  Endocytosis of major histocompatibility complex class I molecules is induced by the HIV–1 Nef protein , 1996, Nature Medicine.

[17]  M. Caron,et al.  Role of β-Arrestin in Mediating Agonist-Promoted G Protein-Coupled Receptor Internalization , 1996, Science.

[18]  J. Benovic,et al.  Beta-arrestin acts as a clathrin adaptor in endocytosis of the beta2-adrenergic receptor. , 1996, Nature.

[19]  G. Milanesi,et al.  NEF-CD 4 Physical Interaction Sensed with the Yeast Two-Hybrid System , 1996 .

[20]  J. S. Sullivan,et al.  Genomic Structure of an Attenuated Quasi Species of HIV-1 from a Blood Transfusion Donor and Recipients , 1995, Science.

[21]  J. Bonifacino,et al.  Interaction of tyrosine-based sorting signals with clathrin-associated proteins. , 1995, Science.

[22]  A. Greenway,et al.  Human immunodeficiency virus type 1 Nef protein inhibits activation pathways in peripheral blood mononuclear cells and T-cell lines , 1995, Journal of virology.

[23]  John L. Sullivan,et al.  Absence of intact nef sequences in a long-term survivor with nonprogressive HIV-1 infection , 1995 .

[24]  J. Skowroński,et al.  Human immunodeficiency virus type 1 Nef and p56lck protein-tyrosine kinase interact with a common element in CD4 cytoplasmic tail. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  B. Goud,et al.  Human immunodeficiency virus type 1 Nef induces accumulation of CD4 in early endosomes , 1995, Journal of virology.

[26]  S. Fields,et al.  Analyzing protein-protein interactions using two-hybrid system. , 1995, Methods in enzymology.

[27]  M. Harris,et al.  Myristoylation-dependent binding of HIV-1 Nef to CD4. , 1994, Journal of molecular biology.

[28]  S. Rhee,et al.  Human immunodeficiency virus type 1 Nef-induced down-modulation of CD4 is due to rapid internalization and degradation of surface CD4 , 1994, Journal of Virology.

[29]  M. Robinson,et al.  The role of clathrin, adaptors and dynamin in endocytosis. , 1994, Current opinion in cell biology.

[30]  M. Lenburg,et al.  Nef induces CD4 endocytosis: Requirement for a critical dileucine motif in the membrane-proximal CD4 cytoplasmic domain , 1994, Cell.

[31]  T. Kirchhausen,et al.  The beta 1 and beta 2 subunits of the AP complexes are the clathrin coat assembly components. , 1993, The EMBO journal.

[32]  S. Elledge,et al.  The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases , 1993, Cell.

[33]  S. Elledge,et al.  The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. , 1993, Genes & development.

[34]  S. Simon,et al.  Endocytosis of ß2 integrins by stimulated human neutrophils analyzed by flow cytometry , 1993, Journal of leukocyte biology.

[35]  R. G. Anderson,et al.  The appendage domain of the AP-2 subunit is not required for assembly or invagination of clathrin-coated pits , 1993, The Journal of cell biology.

[36]  C. Hopkins,et al.  Signal-dependent membrane protein trafficking in the endocytic pathway. , 1993, Annual review of cell biology.

[37]  R. Klausner,et al.  A novel di-leucine motif and a tyrosine-based motif independently mediate lysosomal targeting and endocytosis of CD3 chains , 1992, Cell.

[38]  K. Umesono,et al.  Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors , 1991, Cell.

[39]  R. Desrosiers,et al.  Importance of the nef gene for maintenance of high virus loads and for development of AIDS , 1991, Cell.

[40]  T. Kirchhausen,et al.  Clathrin domains involved in recognition by assembly protein AP-2. , 1991, The Journal of biological chemistry.

[41]  J. Garcia,et al.  Serine phosphorylation-independent downregulation of cell-surface CD4 by nef , 1991, Nature.

[42]  G. Griffiths,et al.  Differential endocytosis of CD4 in lymphocytic and nonlymphocytic cells , 1991, The Journal of experimental medicine.

[43]  E. Tramont,et al.  The human immunodeficiency virus. , 1991, Dermatologic clinics.

[44]  J. Rodman,et al.  Endocytosis and transcytosis. , 1990, Current opinion in cell biology.

[45]  M. Marsh,et al.  Internalization and recycling of CD4 transfected into HeLa and NIH3T3 cells. , 1989, The EMBO journal.

[46]  J. Scheppler,et al.  Down-modulation of MHC-I in a CD4+ T cell line, CEM-E5, after HIV-1 infection. , 1989, Journal of immunology.

[47]  B. Guy,et al.  HIV F/3' orf encodes a phosphorylated GTP-binding protein resembling an oncogene product , 1987, Nature.

[48]  D. Russell,et al.  Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. , 1985, Annual review of cell biology.