A Strategy for O-Glycoproteomics of Enveloped Viruses—the O-Glycoproteome of Herpes Simplex Virus Type 1

PLoS Pathogens

Hiren J. Joshi¹  Hans H. Wandall¹  Ieva Bagdonaite¹  Sergey Y. Vakhrushev¹  Rickard Nordén²  Kristina Nyström²  Sigvard Olofsson²  Sally Dabelsteen³ 

[1] Copenhagen Center for Glycomics, Institute of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen, Denmark;Department of Clinical Virology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden;Institute of Odontology, University of Copenhagen, Copenhagen, Denmark

关键词: Glycosylation;    Herpes simplex virus-1;    Fibroblasts;    Glycoproteins;    Host cells;    Herpesviruses;    Antibodies;    Virus glycoproteins;   

DOI  :  10.1371/journal.ppat.1004784

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Glycosylation of viral envelope proteins is important for infectivity and interaction with host immunity, however, our current knowledge of the functions of glycosylation is largely limited to N-glycosylation because it is difficult to predict and identify site-specific O-glycosylation. Here, we present a novel proteome-wide discovery strategy for O-glycosylation sites on viral envelope proteins using herpes simplex virus type 1 (HSV-1) as a model. We identified 74 O-linked glycosylation sites on 8 out of the 12 HSV-1 envelope proteins. Two of the identified glycosites found in glycoprotein B were previously implicated in virus attachment to immune cells. We show that HSV-1 infection distorts the secretory pathway and that infected cells accumulate glycoproteins with truncated O-glycans, nonetheless retaining the ability to elongate most of the surface glycans. With the use of precise gene editing, we further demonstrate that elongated O-glycans are essential for HSV-1 in human HaCaT keratinocytes, where HSV-1 produced markedly lower viral titers in HaCaT with abrogated O-glycans compared to the isogenic counterpart with normal O-glycans. The roles of O-linked glycosylation for viral entry, formation, secretion, and immune recognition are poorly understood, and the O-glycoproteomics strategy presented here now opens for unbiased discovery on all enveloped viruses.

【 授权许可】

CC BY