title: VP08R from infectious spleen and kidney necrosis virus is a novel component of the virus-mock basement membrane.
abstract: Infectious spleen and kidney necrosis virus (ISKNV), the type species of the genus Megalocytivirus, family Iridoviridae, brings great harm to fish farming. In infected tissues, ISKNV infection is characterized by a unique phenomenon, in that the infected cells are attached by lymphatic endothelial cells (LECs), which are speculated to wall off the infected cells from host immune attack. A viral membrane protein, VP23R, binds and recruits the host nidogen-1 protein to construct a basement membrane (BM)-like structure, termed virus-mock basement membrane (VMBM), on the surface of infected cells to provide attaching sites for LECs. VMBMs do not contain collagen IV protein, which is essential for maintenance of BM integrity and functions. In this study, we identified the VP08R protein encoded by ISKNV. VP08R was predicted to be a secreted protein with a signal peptide but without a transmembrane domain. However, immunofluorescence assays demonstrated that VP08R is located on the plasma membrane of infected cells and shows an expression profile similar to that of VP23R. Coimmunoprecipitation showed that VP08R interacts with both VP23R and nidogen-1, indicating that VP08R is a component of VMBM and is present on the cell membrane by binding to VP23R. Through formation of intermolecular disulfide bonds, VP08R molecules self-organized into a multimer, which may play a role in the maintenance of VMBM integrity and stability. Moreover, the VP08R multimer was easily degraded when the ISKNV-infected cells were lysed, which may be a mechanism for VMBM disassembly when necessary to free LECs and release the mature virions.
  
  
  OBJECTIVE
  Infectious spleen and kidney necrosis virus (ISKNV; genus Megalocytivirus, family Iridovirus) is most harmful to cultured fishes. In tissues, the ISKNV-infected cells are attached by lymphatic endothelial cells (LECs), which are speculated to segregate the host immune system. A viral membrane protein, VP23R, binds and recruits the host nidogen-1 protein to construct virus-mock basement membranes (VMBMs) on the surface of infected cells to provide attaching sites for LECs. Although VMBMs lack the collagen IV network, which is an essential structural part of true BMs, VMBMs still show an intact structure. An ISKNV-encoded VP08R protein can self-assemble into a multimer and bind both VP23R and nidogen-1 to maintain the integrity and stability of VMBMs. On the basis of these facts, we redrew the putative schematic illustration of the VMBM structure. Our study suggests that the virus adopts a strategy to remodel the cellular matrix and may provide an important reference to elucidate BM functions and the mechanisms of lymphangiogenesis.
authors.affiliation: MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, People's Republic of China.
authors.forename: Xiaopeng
authors.initials: X
authors.lastname: Xu
authors.forename: Muting
authors.initials: M
authors.lastname: Yan
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.forename: Ting
authors.initials: T
authors.lastname: Lin
authors.forename: Junliang
authors.initials: J
authors.lastname: Tang
authors.forename: Chaozheng
authors.initials: C
authors.lastname: Li
authors.forename: Shaoping
authors.initials: S
authors.lastname: Weng
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: He
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry*; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D007497:Iridoviridae / Q000235:genetics* / Q000502:physiology*; D008856:Microscopy, Fluorescence; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D015678:Viral Structural Proteins / Q000032:analysis* / Q000235:genetics*
chemical_list: D015678:Viral Structural Proteins