SARS-CoV-2 envelope protein topology in eukaryotic membranes

  1. Gerard Duart
  2. Mª Jesús García-Murria
  3. Brayan Grau
  4. José M. Acosta-Cáceres
  5. Luis Martínez-Gil
  6. Ismael Mingarro

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Abstract

Coronavirus E protein is a small membrane protein found in the virus envelope. Different coronavirus E proteins share striking biochemical and functional similarities, but sequence conservation is limited. In this report, we studied the E protein topology from the new SARS-CoV-2 virus both in microsomal membranes and in mammalian cells. Experimental data reveal that E protein is a single-spanning membrane protein with the N-terminus being translocated across the membrane, while the C-terminus is exposed to the cytoplasmic side (Nt lum /Ct cyt ). The defined membrane protein topology of SARS-CoV-2 E protein may provide a useful framework to understand its interaction with other viral and host components and contribute to establish the basis to tackle the pathogenesis of SARS-CoV-2.

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  1. Version published to 10.1098/rsob.200209
  2. ScreenIT

    SciScore for 10.1101/2020.05.27.118752: (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.

    Table 2: Resources

    Antibodies
    SentencesResources
    Protein glycosylation status was analysed by Western Blot using an anti-c-myc antibody (Sigma), anti-rabbit IgG-peroxidase conjugated (Sigma), and with ECL developing reagent (GE Healthcare).
    anti-c-myc
    suggested: None
    anti-rabbit IgG-peroxidase
    suggested: None
    Software and Algorithms
    SentencesResources
    All oligonucleotides were purchased from Macrogen (Seoul, South Korea).
    Macrogen
    suggested: (Macrogen, RRID:SCR_014454)
    Computer-assisted analysis of E protein sequence: Prediction of transmembrane segments was done using up to 7 of the most common methods available on the Internet: ΔG Predictor [27,28] (http://dgpred.cbr.su.se/), TMHMM [29] (http://www.cbs.dtu.dk/services/TMHMM/), MEMSAT-SVM [30] (http://bioinf.cs.ucl.ac.uk/psipred/), TMpred (https://embnet.vital-it.ch/software/TMPRED_form.html), HMMTop [31] (http://www.enzim.hu/hmmtop/), Phobius [32] (http://phobius.sbc.su.se/) and TOPCONS [33] (http://topcons.net/).
    http://phobius.sbc.su.se/
    suggested: (Phobius, RRID:SCR_015643)

    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 did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    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

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  3. Version published to 10.1101/2020.05.27.118752v1 on bioRxiv