Patterns of human and porcine gammaherpesvirus-encoded BILF1 receptor endocytosis

Mavri, Maša (author), Glisic, Sanja (author), Sencanski, Milan (author), Vrecl, Milka (author), Rosenkilde, Mette Marie (author), Spiess, Katja (author), Kubale, Valentina (author)

Background: The 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 orthologues 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 to EBV-BILF1. Methods: A 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. Results: We 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. Conclusions: The 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.

• Language: English
• Keywords: Herpesvirus 4, human, Dynamins, Caveolins, beta-Arrestins
• Typology: 1.01 - Original Scientific Article
• Organization: VF - Veterinary Faculty
• Publication status: Published
• Publication version: Version of Record
• Publication date: 01.01.2023
• Year: 2023
• Number of pages: Str. 1-19
• Numbering: Vol. 28, [artlice no.] 14
• PID: 20.500.12556/RUL-144509
• UDC: 577
• ISSN on article: 1689-1392
• DOI: 10.1186/s11658-023-00427-y
• COBISS.SI-ID: 141009411
• Publication date in RUL: 27.02.2023

Record is a part of a journal

• Title: Cellular and molecular biology letters. international scientific journal
• Shortened title: Cell. Mol. Biol. Lett.
• Publisher: University of Wrocław
• ISSN: 1689-1392
• COBISS.SI-ID: 3249585

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Projects

• Funder: MESTD - Ministry of Education, Science and Technological Development of Republic of Serbia
• Project number: 451-03-47/2023-01/200017

• Funder: Other - Other funder or multiple funders
• Funding programme: Slovenian-Serbian bilateral project
• Project number: BI-RS/20-21-045