Comprehensive functional analysis of N-linked glycans on ebola virus GP1

Nicholas J. Lennemann, Bethany A. Rhein, Esther Ndungo, Kartik Chandran, Xiangguo Qiu, Wendy Maury

Research output: Contribution to journalArticle

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Abstract

Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. Sitedirected mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. The effects of the loss of GP1 N-linked glycans on Ca2+-dependent (C-type) lectin (CLEC)- dependent transduction were complex, and the effect was unique for each of the CLECs tested. Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. Finally, the removal of all GP1 N-glycans outside the MLD enhanced antiserum and antibody sensitivity. In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency.

Original languageEnglish (US)
Article numbere00862-13
JournalmBio
Volume5
Issue number1
DOIs
StatePublished - Jan 28 2014

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Ebolavirus
Polysaccharides
Mucins
C-Type Lectins
Virus Internalization
Glycoproteins
Glycosylation
Vero Cells
Antibodies
Membrane Glycoproteins
Peritoneal Macrophages
Masks
Mutagenesis
Virion

ASJC Scopus subject areas

  • Microbiology
  • Virology

Cite this

Lennemann, N. J., Rhein, B. A., Ndungo, E., Chandran, K., Qiu, X., & Maury, W. (2014). Comprehensive functional analysis of N-linked glycans on ebola virus GP1. mBio, 5(1), [e00862-13]. https://doi.org/10.1128/mBio.00862-13

Comprehensive functional analysis of N-linked glycans on ebola virus GP1. / Lennemann, Nicholas J.; Rhein, Bethany A.; Ndungo, Esther; Chandran, Kartik; Qiu, Xiangguo; Maury, Wendy.

In: mBio, Vol. 5, No. 1, e00862-13, 28.01.2014.

Research output: Contribution to journalArticle

Lennemann, NJ, Rhein, BA, Ndungo, E, Chandran, K, Qiu, X & Maury, W 2014, 'Comprehensive functional analysis of N-linked glycans on ebola virus GP1', mBio, vol. 5, no. 1, e00862-13. https://doi.org/10.1128/mBio.00862-13
Lennemann, Nicholas J. ; Rhein, Bethany A. ; Ndungo, Esther ; Chandran, Kartik ; Qiu, Xiangguo ; Maury, Wendy. / Comprehensive functional analysis of N-linked glycans on ebola virus GP1. In: mBio. 2014 ; Vol. 5, No. 1.
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