Mechanism for removal of tumor necrosis factor receptor 1 from the cell surface by the adenovirus RIDalpha/beta complex.
Proteins encoded in adenovirus early region 3 have important immunoregulatory properties. We have recently shown that the E3-10.4K/14.5K (RIDalpha/beta) complex downregulates tumor necrosis factor receptor 1 (TNFR1) expression at the plasma membrane. To study the role of the RIDbeta tyrosine sorting motif in the removal of surface TNFR1, tyrosine 122 on RIDbeta was mutated to alanine or phenylalanine. Both RIDbeta mutations not only abolished the downregulation of surface TNFR1 but paradoxically increased surface TNFR1 levels. RID also downregulates other death receptors, such as FAS; however, surface FAS expression was not increased by RIDbeta mutants, suggesting that regulation of TNFR1 and that of FAS by RID are mechanistically different. In the mixing experiments, the wild-type (WT) RID-mediated TNFR1 downregulation was partially inhibited in the presence of RIDbeta mutants, indicating that the mutants compete for TNFR1 access. Indeed, an association between RIDbeta and TNFR1 was shown by coimmunoprecipitation. In contrast, the mutants did not affect the WT RID-induced downregulation of FAS. These differential effects support a model in which RID associates with TNFR1 on the plasma membrane, whereas RID probably associates with FAS in a cytoplasmic compartment. By using small interfering RNA against the mu2 subunit of adaptor protein 2, dominant negative dynamin construct K44A, and the lysosomotropic agents bafilomycin A1 and ammonium chloride, we also demonstrated that surface TNFR1 was internalized by RID by a clathrin-dependent process involving mu2 and dynamin, followed by degradation of TNFR1 via an endosomal/lysosomal pathway.
Chin
Y Rebecca
YR
Department of Microbiology and Immunology, Forchheimer Building, Room 411, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461, USA. ychin@aecom.yu.edu
Horwitz
Marshall S
MS
D048868:Adaptor Proteins, Signal Transducing; D017503:Adenovirus E3 Proteins / Q000235:genetics / Q000502:physiology*; D000260:Adenoviruses, Human / Q000502:physiology*; D017209:Apoptosis; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D056970:Co-Repressor Proteins; D015536:Down-Regulation; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*; D047888:Receptors, Tumor Necrosis Factor, Type I; D053319:Tumor Necrosis Factor Decoy Receptors; D014779:Virus Replication
D048868:Adaptor Proteins, Signal Transducing; D017503:Adenovirus E3 Proteins; D002352:Carrier Proteins; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D053319:Tumor Necrosis Factor Decoy Receptors; C069521:recombinant human tumor necrosis factor-binding protein-1