Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Biomolecular Fluorescence Complementation Profiling and Artificial Intelligence Structure Prediction of the Kaposi’s Sarcoma-associated Herpesvirus ORF18 and ORF30 Interaction

Version 1 : Received: 25 June 2022 / Approved: 27 June 2022 / Online: 27 June 2022 (10:15:34 CEST)

How to cite: Maeda, Y.; Watanabe, T.; Izumi, T.; Kuriyama, K.; Ohno, S.; Fujimuro, M. Biomolecular Fluorescence Complementation Profiling and Artificial Intelligence Structure Prediction of the Kaposi’s Sarcoma-associated Herpesvirus ORF18 and ORF30 Interaction . Preprints 2022, 2022060359 (doi: 10.20944/preprints202206.0359.v1). Maeda, Y.; Watanabe, T.; Izumi, T.; Kuriyama, K.; Ohno, S.; Fujimuro, M. Biomolecular Fluorescence Complementation Profiling and Artificial Intelligence Structure Prediction of the Kaposi’s Sarcoma-associated Herpesvirus ORF18 and ORF30 Interaction . Preprints 2022, 2022060359 (doi: 10.20944/preprints202206.0359.v1).

Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi’s sarcoma, primary effusion lymphoma (PEL) and multicentric Castleman’s disease. During KSHV lytic infection, lytic-related genes, categorized as immediate-early, early, and late genes, are expressed in a temporal manner. The transcription of late genes requires the virus-specific pre-initiation complex (vPIC), which consists of viral transcription factors. However, the characterization of the protein-protein interactions of the vPIC factors has not been completely elucidated. KSHV ORF18 is one of the vPIC factors, and its interaction with other viral factors has not been sufficiently revealed. Here, we analyzed the interaction between ORF18 and another vPIC factor, ORF30, in living cells using the bimolecular fluorescence complementation (BiFC) assay. We identified four amino-acid residues (Leu29, Gln36, His41 and Trp170) on ORF18 that were responsible for its interaction with ORF30. The artificial intelligence (AI) system AlphaFold2 predicted that identified four residues are exposed to the surface of ORF18 and located in proximity to each other in the surface of ORF18. Thus, AI-predicted model supports the importance of four residues for binding of ORF18 to ORF30. Our results indicated that wet experiments in combination with AI may enhance the structural characterization of vPIC protein-protein interactions.

Keywords

Kaposi’s sarcoma-associated virus (KSHV); Viral pre-initiation complex (vPIC); Bimolecular fluorescence complementation (BiFC); Artificial intelligence (AI) structure prediction; AlphaFold2

Subject

MEDICINE & PHARMACOLOGY, Pathology & Pathobiology

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