title: Constitutive activation of T cells by γ2-herpesviral GPCR through the interaction with cellular CXCR4.
abstract: Members of the herpesviral family use multiple strategies to hijack infected host cells and exploit cellular signaling for their pathogenesis and latent infection. Among the most intriguing weapons in the arsenal of pathogenic herpesviruses are the constitutively active virally-encoded G protein-coupled receptors (vGPCRs). Even though vGPCRs contribute to viral pathogenesis such as immune evasion and proliferative disorders, the molecular details of how vGPCRs continuously activate cellular signaling are largely unknown. Here, we report that the vGPCR of Herpesvirus saimiri (HVS), an oncogenic γ2-herpesvirus, constitutively activates T cells via a heteromeric interaction with cellular CXCR4. Constitutive T cell activation also occurs with expression of the vGPCR of Kaposi's sarcoma-associated herpesvirus (KSHV), but not the vGPCR of Epstein-Barr virus. Expression of HVS vGPCR down-regulated the surface expression of CXCR4 but did not induce the degradation of the chemokine receptor, suggesting that vGPCR/CXCR4 signaling continues in cytosolic compartments. The physical association of vGPCR with CXCR4 was demonstrated by proximity ligation assay as well as immunoprecipitation. Interestingly, the constitutive activation of T cells by HVS vGPCR is independent of proximal T cell receptor (TCR) signaling molecules, such as TCRβ, Lck, and ZAP70, whereas CXCR4 silencing by shRNA abolished T cell activation by vGPCRs of HVS and KSHV. Furthermore, previously identified inactive vGPCR mutants failed to interact with CXCR4. These findings on the positive cooperativity of vGPCR with cellular CXCR4 in T cell activation extend our current understanding of the molecular mechanisms of vGPCR function and highlight the importance of heteromerization for GPCR activity.
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Eun-Kyung
authors.initials: EK
authors.lastname: Kwon
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Chan-Ki
authors.initials: CK
authors.lastname: Min
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Jae-Won
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Abdimadiyeva
authors.initials: A
authors.lastname: Aigerim
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Department of Life Sciences, Pohang University of Science and Technology, Pohang 37673, Republic of Korea.
authors.forename: Hyun-Jun
authors.initials: HJ
authors.lastname: Nam
authors.affiliation: Department of Life Sciences, Pohang University of Science and Technology, Pohang 37673, Republic of Korea.
authors.forename: Seong Kyu
authors.initials: SK
authors.lastname: Han
authors.affiliation: Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea.
authors.forename: Kuglae
authors.initials: K
authors.lastname: Kim
authors.affiliation: Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea.
authors.forename: Jeong Seok
authors.initials: JS
authors.lastname: Cha
authors.affiliation: Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea.
authors.forename: Hoyoung
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Life Sciences, Pohang University of Science and Technology, Pohang 37673, Republic of Korea.
authors.forename: Sanguk
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea.
authors.forename: Hyun-Soo
authors.initials: HS
authors.lastname: Cho
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Myung-Sik
authors.initials: MS
authors.lastname: Choi
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Institute of Endemic Disease, Seoul National University Medical Research Center and Bundang Hospital, Seoul 03080, Republic of Korea. Electronic address: chonh@snu.ac.kr.
authors.forename: Nam-Hyuk
authors.initials: NH
authors.lastname: Cho
mesh_terms: D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D061251:Primary Cell Culture; D011485:Protein Binding; D055503:Protein Multimerization; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D019707:Receptors, Chemokine / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: C527276:CXCR4 protein, human; C115448:G protein-coupled receptor, Human herpesvirus 8; D016693:Receptors, Antigen, T-Cell, alpha-beta; D019718:Receptors, CXCR4; D019707:Receptors, Chemokine; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human
keywords: CXCR4; T cells; Viral G protein-coupled receptor; γ-Herpesvirus