Patterns of human and porcine gammaherpesvirus-encoded BILF1 receptor endocytosis

Cellular & Molecular Biology Letters

Research Letter

Sanja Glišić¹  Milan Senćanski¹  Mette M. Rosenkilde²  Katja Spiess³  Maša Mavri⁴  Milka Vrecl⁴  Valentina Kubale⁴ 

[1] Center for Multidisciplinary Research, Institute of Nuclear Sciences VINCA, University of Belgrade, Belgrade, Serbia;Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark;Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark;Department of Virus and Microbiological Special Diagnostics, Statens Serum Institute, Copenhagen, Denmark;Institute for preclinical sciences, Veterinary Faculty, Ljubljana, Slovenia;

关键词: vGPCR;    BILF1;    Endocytosis;    Internalization;    Dynamin;    Caveolin;    β-Arrestin;    EBV;    PLHV1-2;   

DOI  :  10.1186/s11658-023-00427-y

 received in 2022-10-24, accepted in 2023-01-30,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundThe viral G-protein-coupled receptor (vGPCR) BILF1 encoded by the Epstein–Barr virus (EBV) is an oncogene and immunoevasin and can downregulate MHC-I molecules at the surface of infected cells. MHC-I downregulation, which presumably occurs through co-internalization with EBV-BILF1, is preserved among BILF1 receptors, including the three BILF1 orthologs encoded by porcine lymphotropic herpesviruses (PLHV BILFs). This study aimed to understand the detailed mechanisms of BILF1 receptor constitutive internalization, to explore the translational potential of PLHV BILFs compared with EBV-BILF1.MethodsA novel real-time fluorescence resonance energy transfer (FRET)-based internalization assay combined with dominant-negative variants of dynamin-1 (Dyn K44A) and the chemical clathrin inhibitor Pitstop2 in HEK-293A cells was used to study the effect of specific endocytic proteins on BILF1 internalization. Bioluminescence resonance energy transfer (BRET)-saturation analysis was used to study BILF1 receptor interaction with β-arrestin2 and Rab7. In addition, a bioinformatics approach informational spectrum method (ISM) was used to investigate the interaction affinity of BILF1 receptors with β-arrestin2, AP-2, and caveolin-1.ResultsWe identified dynamin-dependent, clathrin-mediated constitutive endocytosis for all BILF1 receptors. The observed interaction affinity between BILF1 receptors and caveolin-1 and the decreased internalization in the presence of a dominant-negative variant of caveolin-1 (Cav S80E) indicated the involvement of caveolin-1 in BILF1 trafficking. Furthermore, after BILF1 internalization from the plasma membrane, both the recycling and degradation pathways are proposed for BILF1 receptors.ConclusionsThe similarity in the internalization mechanisms observed for EBV-BILF1 and PLHV1-2 BILF1 provide a foundation for further studies exploring a possible translational potential for PLHVs, as proposed previously, and provides new information about receptor trafficking.Graphical Abstract

【 授权许可】

CC BY   
© The Author(s) 2023

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