Protein N -glycosylation is essential for SARS-CoV-2 infection

  1. Aitor Casas-Sanchez
  2. Alessandra Romero-Ramirez
  3. Eleanor Hargreaves
  4. Cameron C. Ellis
  5. Brian I. Grajeda
  6. Igor Estevao
  7. Edward I. Patterson
  8. Grant L. Hughes
  9. Igor C. Almeida
  10. Tobias Zech
  11. Álvaro Acosta-Serrano

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

SARS-CoV-2 extensively N -glycosylates its spike proteins, which are necessary for host cell invasion and the target of both vaccines and immunotherapies. These sugars are predicted to help mediate spike binding to the host receptor by stabilizing its ‘open’ conformation and evading host immunity. Here, we investigated both the essentiality of the host N -glycosylation pathway and SARS-CoV-2 N -glycans for infection. Inhibition of host N -glycosylation using RNAi or FDA-approved drugs reduced virus infectivity, including that of several variants. Under these conditions, cells produced less virions and some completely lost their infectivity. Furthermore, partial deglycosylation of intact virions showed that surface-exposed N -glycans are critical for cell invasion. Altogether, spike N -glycosylation is a targetable pathway with clinical potential for treatment or prevention of COVID-19.

Article activity feed

  3. Version published to 10.1101/2021.02.05.429940v2 on bioRxiv
  4. ScreenIT

    SciScore for 10.1101/2021.02.05.429940: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Primary antibodies used: anti-STT3A (HPA030735, Cambridge Bioscience), anti-STT3B (A15574, Universal Biologicals), anti-GANAB (A13851, Universal Biologicals), anti-MGAT1 (CSB-PA013773ESR1Hu, 2B Scientific) and anti-GAPDH (10494-1-AP, ProteinTech).
    anti-STT3A (HPA030735
    suggested: (Atlas Antibodies Cat# HPA030735, RRID:AB_10603342)
    anti-STT3B
    suggested: (ABclonal Cat# A15574, RRID:AB_2762978)
    anti-GANAB
    suggested: (ABclonal Cat# A13851, RRID:AB_2760703)
    anti-MGAT1
    suggested: None
    CSB-PA013773ESR1Hu
    suggested: None
    anti-GAPDH
    suggested: (Proteintech Cat# 10494-1-AP, RRID:AB_2263076)
    Blots were then washed 3 times under agitation for 5 minutes in TBS-T before incubation with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG secondary antibody (Cell Signalling) for 1 hour at RT.
    anti-rabbit IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK293ACE-2 cells (lentiviral expression of human angiotensin-converting enzyme 2, source) were cultured in DMEM supplemented with 10% FBS, 1mM sodium pyruvate (Gibco) and 2 μg/mL puromycin (Invivogen) at 37°C and 5% CO2.
    HEK293ACE-2
    suggested: None
    Plaque assay: Vero E6 were cultured in DMEM 10% FBS in 24-well plates and allowed to reach confluency overnight.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Images were analyzed using Fiji (ImageJ) by thresholding to select positive signals only and measuring area coverage of DAPI channel (total number of cells) and mean intensity of the Alexa Fluor channel (infected cells).
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 10. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2021.02.05.429940v1 on bioRxiv