The E3 Ubiquitin Ligase RNF5 Facilitates SARS-CoV-2 Membrane Protein-Mediated Virion Release

  1. Zhen Yuan
  2. Bing Hu
  3. Hurong Xiao
  4. Xuan Tan
  5. Yan Li
  6. Ke Tang
  7. Yonghui Zhang
  8. Kun Cai
  9. Binbin Ding

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Abstract

Enveloped virus particles are released from the membranes of host cells, and viral membrane proteins (M) are critical for this process. A better understanding of the molecular mechanisms of SARS-CoV-2 assembly and budding is critical for the development of antiviral therapies.

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  1. Version published to 10.1128/mbio.03168-21
  2. ScreenIT

    SciScore for 10.1101/2021.02.28.433287: (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
    Antibodies and Reagents: Mouse monoclonal anti-FLAG (F1804), anti-HA (H3663) and ERGIC53 (SAB2101351) were obtained from Sigma-Aldrich.
    anti-FLAG
    suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)
    F1804
    suggested: (Sigma-Aldrich Cat# F1804, RRID:AB_262044)
    anti-HA
    suggested: (Sigma-Aldrich Cat# H3663, RRID:AB_262051)
    H3663
    suggested: None
    ERGIC53
    suggested: (Sigma-Aldrich Cat# SAB2101351, RRID:AB_10604834)
    Anti-Tubulin (E7-S) was obtained from Developmental Studies Hybridoma Bank.
    Anti-Tubulin
    suggested: None
    Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 568 conjugate (A11031), goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 568 conjugate (A11036), goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (A32723), goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (A32731) and goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor® 405 (A31553) were obtained from Thermo Fisher Scientific.
    anti-Rabbit IgG
    suggested: (Thermo Fisher Scientific Cat# A-11036, RRID:AB_10563566)
    A32731
    suggested: (Thermo Fisher Scientific Cat# A32731, RRID:AB_2633280)
    anti-Mouse IgG
    suggested: (Molecular Probes Cat# A-31553, RRID:AB_221604)
    A31553
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    rnf5 KO HEK293T, AD293, HeLa and Vero cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; GIBCO) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) at 37°C with 5% CO2.
    HEK293T
    suggested: None
    AD293
    suggested: NCBI_Iran Cat# C559, RRID:CVCL_9804)
    HeLa
    suggested: None
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    The significance of the variability between different groups was determined by two-way analyses of variance using GraphPad Prism software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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: 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

    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.

  3. Version published to 10.1101/2021.02.28.433287 on bioRxiv