Two Distinct Transport Motifs in the Adenovirus E3/10.4-14.5 Proteins Act in Concert to Down-modulate Apoptosis Receptors and the Epidermal Growth Factor Receptor

Hilgendorf, Annette; Lindberg, Johan; Ruzsics, Zsolt; Höning, Stefan; Elsing, A.; Löfqvist, Madelaine; Engelmann, Hartmut; Burgert, Hans‐Gerhard

doi:10.1074/jbc.m310038200

Cited by 33 publications

(58 citation statements)

References 66 publications

(73 reference statements)

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“…1(b)]. These results agree with previous observations that the tyrosine sorting motif of RIDb plays an important role in determining surface levels of RID (Hilgendorf et al, 2003) and TNFR1 (Chin & Horwitz, 2005). Interestingly, although there was less RIDb(YF) mutant present intracellularly than WT RIDb, similar amounts of intracellular TNFR1 were co-immunoprecipitated [middle panel of Fig.…”

supporting

confidence: 82%

“…These data suggested that, instead of accelerating the internalization of TNFR1, RID and clathrin play an important role in Experiments in this study did not address whether RID was degraded together with TNFR1 in the lysosomes. However, previous observations showed limited co-localization of RIDb and FAS in lysosomes (Hilgendorf et al, 2003). In addition, whereas EGFR was degraded in lysosomes, RIDa was retained on the limiting membrane of multivesicular bodies (Crooks et al, 2000).…”

mentioning

confidence: 76%

“…However, previous observations showed limited co-localization of RIDb and FAS in lysosomes (Hilgendorf et al, 2003). In addition, whereas EGFR was degraded in lysosomes, RIDa was retained on the limiting membrane of multivesicular bodies (Crooks et al, 2000).…”

mentioning

confidence: 76%

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Adenovirus RID complex enhances degradation of internalized tumour necrosis factor receptor 1 without affecting its rate of endocytosis

Chin

Horwitz

2006

Journal of General Virology

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The receptor internalization and degradation (RID) complex of adenovirus plays an important role in modulating the immune response by downregulating the surface levels of tumour necrosis factor receptor 1 (TNFR1), thereby inhibiting NF-kB activation. Total cellular content of TNFR1 is also reduced in the presence of RID, which can be inhibited by treatment with lysosomotropic agents. In this report, surface biotinylation experiments revealed that, although RID and TNFR1 were able to form a complex on the cell surface, the rate of TNFR1 endocytosis was not affected by RID. However, the degradation of internalized TNFR1 was enhanced significantly in the presence of RID. Therefore, these data suggest that RID downregulates TNFR1 levels by altering the fate of internalized TNFR1 that becomes associated with RID at the plasma membrane, probably by promoting its sorting into endosomal/lysosomal degradation compartments.The successful replication and survival of viruses in the host requires evasion of the immune system. Many of the immunomodulatory genes of adenovirus (Ad) are encoded in early region 3 (E3) (Fessler et al., 2004b;Horwitz, 2004). The E3 receptor internalization and degradation (RID) complex, composed of two RIDa and one RIDb subunits, downregulates a specific set of plasma-membrane receptors, including FAS (Shisler et al., 1997;Elsing & Burgert, 1998;Tollefson et al., 1998), tumour necrosis factor (TNF)-related apoptosis-inducing ligand receptor 1 (TRAIL-R1) (Benedict et al., 2001;Tollefson et al., 2001) and epidermal growth factor receptor (EGFR) (Carlin et al., 1989;Tollefson et al., 1991). Together with another E3 protein, E3/6.7K, RID also reduces the surface expression of TRAIL-R2 (Lichtenstein et al., 2004). Whilst both of the RID subunits are critical for the downregulation of FAS and TRAIL receptors, it appears that the RIDa subunit is sufficient to downregulate EGFR under certain experimental conditions (Hoffman et al., 1990). Recently, we demonstrated that RID also downregulates TNFR1, thereby inhibiting TNF-induced NF-kB activation, which mediates the transcription of a large number of genes involved in inflammation and immune responses such as chemokine expression (Fessler et al., 2004a;Delgado-Lopez & Horwitz, 2006).In vivo experiments have shown that RID is an essential component of the Ad E3 cassette, which facilitates transplantation of allogeneic pancreatic cells (Efrat et al., 1995) and decreases autoimmune type 1 diabetes incidence (von Herrath et al., 1997;Efrat et al., 2001;Pierce et al., 2003). To potentially use RID as a therapeutic immunomodulator, it is important to elucidate the molecular mechanism of RIDmediated downregulation of receptors. We recently showed that the tyrosine sorting motifs of RIDb and clathrin play important roles in the downregulation of TNFR1, and that RID-mediated TNFR1 degradation occurs via an endosomal/ lysosomal pathway (Chin & Horwitz, 2005). In this report, we further examined the effect of RID on TNFR1 trafficking. Specifically, we tested the abili...

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supporting

confidence: 82%

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confidence: 76%

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Adenovirus RID complex enhances degradation of internalized tumour necrosis factor receptor 1 without affecting its rate of endocytosis

Chin

Horwitz

2006

Journal of General Virology

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“…For silver staining of polyacrylamide gels a commercial kit (BIORAD) was used. Proteins were blotted as described previously (49). E3/49K was detected with either R48 or mAb 4D1.…”

Section: Methodsmentioning

confidence: 99%

A unique secreted adenovirus E3 protein binds to the leukocyte common antigen CD45 and modulates leukocyte functions

Windheim

Southcombe

Kremmer

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Significance Human adenoviruses encode Early region 3 (E3) proteins that manipulate the host immune response to establish an infection or to persist longer. To date, only a few E3 functions from a single adenovirus species (C) have been characterized, all of which act directly on infected cells. Here we describe a secreted E3 protein that is uniquely expressed by species D adenoviruses. This protein targets noninfected leukocytes using a cell surface phosphatase as a receptor. We provide evidence that this interaction suppresses leukocyte activation and effector functions, implying that species D adenoviruses can affect the host distant from the site of infection.

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“…Cell lines expressing E3/19K mutants E5, P9, T14, K27, I37, K42, W52, G55, Y60, V62, V64, M82, M87, W96, and P97 were established by transfection of 293 cells with mutagenized Ad2 EcoRI D fragments. Culture of 293 cells, transfection, and subsequent selection was performed as described (30,62).…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Conserved Amino Acids within the Adenovirus 2 E3/19K Protein Differentially Affect Downregulation of MHC Class I and MICA/B Proteins

Sester

Koebernick

Owen

et al. 2009

The Journal of Immunology

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Successful establishment and persistence of adenovirus (Ad) infections are facilitated by immunosubversive functions encoded in the early transcription unit 3 (E3). The E3/19K protein has a dual role, preventing cell surface transport of MHC class I/HLA class I (MHC-I/HLA-I) Ags and the MHC-I–like molecules (MHC-I chain-related chain A and B [MICA/B]), thereby inhibiting both recognition by CD8 T cells and NK cells. Although some crucial functional elements in E3/19K have been identified, a systematic analysis of the functional importance of individual amino acids is missing. We now have substituted alanine for each of 21 aas in the luminal domain of Ad2 E3/19K conserved among Ads and investigated the effects on HLA-I downregulation by coimmunoprecipitation, pulse-chase analysis, and/or flow cytometry. Potential structural alterations were monitored using conformation-dependent E3/19K-specific mAbs. The results revealed that only a small number of mutations abrogated HLA-I complex formation (e.g., substitutions W52, M87, and W96). Mutants M87 and W96 were particularly interesting as they exhibited only minimal structural changes suggesting that these amino acids make direct contacts with HLA-I. The considerable number of substitutions with little functional defects implied that E3/19K may have additional cellular target molecules. Indeed, when assessing MICA/B cell-surface expression we found that mutation of T14 and M82 selectively compromised MICA/B downregulation with essentially no effect on HLA-I modulation. In general, downregulation of HLA-I was more severely affected than that of MICA/B; for example, substitutions W52, M87, and W96 essentially abrogated HLA-I modulation while largely retaining the ability to sequester MICA/B. Thus, distinct conserved amino acids seem preferentially important for a particular functional activity of E3/19K.

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Two Distinct Transport Motifs in the Adenovirus E3/10.4-14.5 Proteins Act in Concert to Down-modulate Apoptosis Receptors and the Epidermal Growth Factor Receptor

Cited by 33 publications

References 66 publications

Adenovirus RID complex enhances degradation of internalized tumour necrosis factor receptor 1 without affecting its rate of endocytosis

Adenovirus RID complex enhances degradation of internalized tumour necrosis factor receptor 1 without affecting its rate of endocytosis

A unique secreted adenovirus E3 protein binds to the leukocyte common antigen CD45 and modulates leukocyte functions

Conserved Amino Acids within the Adenovirus 2 E3/19K Protein Differentially Affect Downregulation of MHC Class I and MICA/B Proteins

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