Characterization of Host Cell Potential Proteins Interacting with OsHV-1 Membrane Proteins.

Jiangnan Yu,Changming Bai,Bowen Huang,Chongming Wang,Ying Liu,Dandan Wang,Mengli Yao,Chen Li,Lusheng Xin

doi:10.3390/v13122518

Abstract

The interaction between viral membrane associate proteins and host cellular surface molecules should facilitate the attachment and entry of OsHV-1 into host cells. Thus, blocking the putative membrane proteins ORF25 and ORF72 of OsHV-1 with antibodies that have previously been reported to subdue OsHV-1 replication in host cells, especially ORF25. In this study, prey proteins in host hemocytes were screened by pull-down assay with recombinant baits ORF25 and ORF72, respectively. Gene Ontology (GO) analysis of these prey proteins revealed that most of them were mainly associated with binding, structural molecule activity and transport activity in the molecular function category. The protein–protein interaction (PPI) network of the prey proteins was constructed by STRING and clustered via K-means. For both ORF25 and ORF72, three clusters of these prey proteins were distinguished that were mainly associated with cytoskeleton assembly, energy metabolism and nucleic acid processing. ORF25 tended to function in synergy with actins, while ORF72 functioned mainly with tubulins. The above results suggest that these two putative membrane proteins, ORF25 and ORF72, might serve a role in the transport of viral particles with the aid of a cytoskeleton inside cells.

Highlights

Herpesviruses can infect a broad range of hosts both on land and in the sea, and they have evolved successful approaches to infect different cell types, starting with virus attachment and entry into target cells [1]
Lane M, protein marker; lane 1, free Ni2+ Sepharose beads; lane 2, Ni2+ Sepharose beads binding with rORF72; lane 3, Ni2+ Sepharose beads binding with rORF25; lane 4, the input of hemocyte lysates, a–f represents specific bands in lane 2 and 3
Further investigation into how host cell proteins interacted with ORF25 and ORF72 was carried out by pull-down assay, revealing how these two viral proteins might function in an Ostreid herpesvirus 1 (OsHV-1) infection

Summary

Introduction

Herpesviruses can infect a broad range of hosts both on land and in the sea, and they have evolved successful approaches to infect different cell types, starting with virus attachment and entry into target cells [1]. In contrast to small enveloped viruses that encode one or two membrane glycoproteins to mediate their entry into specific cells, herpesviruses are equipped with more than a dozen membrane glycoproteins, which endows them with more opportunities and options when ascertaining successful entry according to cell types [2]. The specific binding between glycoproteins and cell corresponding entry receptors triggers gB to execute membrane fusion, such as the binding between gD of HSV-1 (herpes simplex viruse-1) and host cellular membrane nectin-1 [4], and the binding between gp of EBV (Epstein–Barr virus) and human leukocyte antigen (HLA) class II molecules on B lymphocytes [5]

Methods

Results

Discussion

Conclusion