= Emerging. More than 5 years before clinical availability.
= Expected to be clinically available in 1 to 4 years.
= Clinically available now.
MSACL 2018 EU : Bagdonaite

MSACL 2018 EU Abstract

Topic: Glycomics

Global Glycoproteomic Analysis of O-glycosylation in Human Herpesviruses

Ieva Bagdonaite (Presenter)
University of Copenhagen

Authors: Ieva Bagdonaite (1) , Rickard Nordén (2), Hiren J. Joshi (1), Sarah L. King (1), Sergey Y. Vakhrushev (1), Sigvard Olofsson (2), and Hans H. Wandall (1)
(1) Copenhagen Center for Glycomics, University of Copenhagen, Copenhagen, Denmark (2) Department of Infectious Diseases, University of Gothenburg, Gothenburg, Sweden

Short Abstract

Information on site-specific O-glycosylation of viral envelope glycoproteins is generally very limited despite important functions. We present a powerful mass-spectrometry based strategy to globally identify O-glycosylation sites on viral envelope proteins of a given virus in the context of a productive infection. We successfully utilized the strategy to map O-linked glycosylation sites on several complex herpesviruses demonstrating that O-glycosylation is widely distributed on most envelope proteins. Moreover, we used genetically engineered keratinocytes lacking O-glycan elongation capacity to demonstrate that O-linked glycans are indeed important for HSV-1 biology as HSV-1 particles produced in these cells had significantly lower titers compared to wild-type keratinocytes. These tools enable wider discovery and detailed analysis of the role of site-specific O-glycosylation in virology.

Long Abstract

Introduction

Herpesviruses are among the most complex and widespread human viruses and cause a number of diseases ranging from cold sores to genital infections and encephalitis. Virally encoded envelope glycoproteins serve as mediators of cell entry as well as modulators of the immune response and are attractive vaccine targets. However, only few herpesvirus-targeted vaccines have proven to be effective despite extensive efforts. While the structure and function of viral envelope proteins are well known, limited information exists on their potential modification by the host’s glycosylation apparatus.

Methods

We have recently established a “bottom-up” mass spectrometry-based technique for mapping viral O-glycosylation sites using Herpes simplex virus type 1-infected human fibroblasts as a model. In addition, we used glycoengineered keratinocytes to assess the biological impact of loss of O-glycan elongation.

Results

Here we applied our proteome-wide discovery platform for mapping mucin-type O-glycosylation sites for several other clinically significant members of the Herpesvirus family – Herpes simplex virus type 2 (HSV-2), Varicella zoster virus (VZV), human cytomegalovirus (HCMV), and Epstein-Barr virus (EBV). We also show that the technique is applicable to various samples, including infected cell lysates and clinical material. In addition, we have demonstrated that O-linked glycan elongation is important for generation of progeny virus and early immune sensing.

Conclusions & Discussion

Given the potential importance of O-linked glycans in virus entry, formation, secretion and recognition by host’s immune system, these findings will serve as important tools for future studies on herpesvirus biology and vaccine development.


References & Acknowledgements:

1. Bagdonaite I, Norden R, Joshi HJ, Dabelsteen S, Nystrom K, Vakhrushev SY, et al. A strategy for O-glycoproteomics of enveloped viruses--the O-glycoproteome of herpes simplex virus type 1. PLoS pathogens. 2015;11(4):e1004784.

2. Bagdonaite I, Norden R, Joshi HJ, King SL, Vakhrushev SY, Olofsson S, et al. Global Mapping of O-Glycosylation of Varicella Zoster Virus, Human Cytomegalovirus, and Epstein-Barr Virus. J Biol Chem. 2016;291(23):12014-28.

3. Iversen MB, Reinert LS, Thomsen MK, Bagdonaite I, Nandakumar R, Cheshenko N, et al. An innate antiviral pathway acting before interferons at epithelial surfaces. Nature immunology. 2016;17(2):150-8.

4. Bagdonaite I, Wandall HH. Global aspects of viral glycosylation. Glycobiology. 2018;28(7):443-67.


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