title: Ovine Herpesvirus 2 Glycoproteins B, H, and L Are Sufficient for, and Viral Glycoprotein Ov8 Can Enhance, Cell-Cell Membrane Fusion.
abstract: Ovine herpesvirus 2 (OvHV-2) is a gammaherpesvirus in the genusMacavirusthat is carried asymptomatically by sheep. Infection of poorly adapted animals with OvHV-2 results in sheep-associated malignant catarrhal fever, a fatal disease characterized by lymphoproliferation and vasculitis. There is no treatment or vaccine for the disease and no cell culture system to propagate the virus. The lack of cell culture has hindered studies of OvHV-2 biology, including its entry mechanism. As an alternative method to study OvHV-2 glycoproteins responsible for membrane fusion as a part of the entry mechanism, we developed a virus-free cell-to-cell membrane fusion assay to identify the minimum required OvHV-2 glycoproteins to induce membrane fusion. OvHV-2 glycoproteins B, H, and L (gB, gH, and gL) were able to induce membrane fusion together but not when expressed individually. Additionally, open reading frame Ov8, unique to OvHV-2, was found to encode a transmembrane glycoprotein that can significantly enhance membrane fusion. Thus, OvHV-2 gB, gH, and gL are sufficient to induce membrane fusion, while glycoprotein Ov8 plays an enhancing role by an unknown mechanism.IMPORTANCEHerpesviruses enter cells via attachment of the virion to the cellular surface and fusion of the viral envelope with cellular membranes. Virus-cell membrane fusion is an important step for a successful viral infection. Elucidating the roles of viral glycoproteins responsible for membrane fusion is critical toward understanding viral entry. Entry of ovine herpesvirus 2 (OvHV-2), the causative agent of sheep associated-malignant catarrhal fever, which is one of the leading causes of death in bison and other ungulates, has not been well studied due to the lack of a cell culture system to propagate the virus. The identification of OvHV-2 glycoproteins that mediate membrane fusion may help identify viral and/or cellular factors involved in OvHV-2 cell tropism and will advance investigation of cellular factors necessary for virus-cell membrane fusion. We found that OvHV-2 glycoproteins B, H, and L are sufficient for, and viral glycoprotein Ov8 can significantly enhance, cell-cell membrane fusion.
authors.affiliation: Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA.
authors.forename: Salim M
authors.initials: SM
authors.lastname: AlHajri
authors.affiliation: Animal Disease Research Unit, USDA-Agricultural Research Service, Washington State University, Pullman, Washington, USA.|Paul G. Allen School for Global Animal Health, Washington State University, Pullman, Washington, USA.
authors.forename: Cristina W
authors.identifier: http://orcid.org/0000-0002-8168-5448
authors.initials: CW
authors.lastname: Cunha
authors.affiliation: Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA.|Paul G. Allen School for Global Animal Health, Washington State University, Pullman, Washington, USA.
authors.forename: Anthony V
authors.initials: AV
authors.lastname: Nicola
authors.affiliation: Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA.|Paul G. Allen School for Global Animal Health, Washington State University, Pullman, Washington, USA.
authors.forename: Hector C
authors.initials: HC
authors.lastname: Aguilar
authors.affiliation: Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA.|Animal Disease Research Unit, USDA-Agricultural Research Service, Washington State University, Pullman, Washington, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington, USA tausns@vetmed.wsu.edu.|Animal Disease Research Unit, USDA-Agricultural Research Service, Washington State University, Pullman, Washington, USA.
authors.forename: Naomi S
authors.initials: NS
authors.lastname: Taus
mesh_terms: D000818:Animals; D002459:Cell Fusion; D002460:Cell Line; D018142:Gammaherpesvirinae / Q000502:physiology*; D054884:Host-Pathogen Interactions; D015678:Viral Structural Proteins / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D015678:Viral Structural Proteins
keywords: glycoproteins; membrane fusion; ovine herpesvirus 2