Broad and differential animal ACE2 receptor usage by SARS-CoV-2

  1. Xuesen Zhao
  2. Danying Chen
  3. Robert Szabla
  4. Mei Zheng
  5. Guoli Li
  6. Pengcheng Du
  7. Shuangli Zheng
  8. Xinglin Li
  9. Chuan Song
  10. Rui Li
  11. Ju-Tao Guo
  12. Murray Junop
  13. Hui Zeng
  14. Hanxin Lin

Reviewed by

Read the full article See related articles

Discuss this preprint

Start a discussion What are Sciety discussions?

Listed in

Log in to save this article

Abstract

The COVID-19 pandemic has caused an unprecedented global public health and economy crisis. The origin and emergence of its causal agent, SARS-CoV-2, in the human population remains mysterious, although bat and pangolin were proposed to be the natural reservoirs. Strikingly, comparing to the SARS-CoV-2-like CoVs identified in bats and pangolins, SARS-CoV-2 harbors a polybasic furin cleavage site in its spike (S) glycoprotein. SARS-CoV-2 uses human ACE2 as its receptor to infect cells. Receptor recognition by the S protein is the major determinant of host range, tissue tropism, and pathogenesis of coronaviruses. In an effort to search for the potential intermediate or amplifying animal hosts of SARS-CoV-2, we examined receptor activity of ACE2 from 14 mammal species and found that ACE2 from multiple species can support the infectious entry of lentiviral particles pseudotyped with the wild-type or furin cleavage site deficient S protein of SARS-CoV-2. ACE2 of human/rhesus monkey and rat/mouse exhibited the highest and lowest receptor activity, respectively. Among the remaining species, ACE2 from rabbit and pangolin strongly bound to the S1 subunit of SARS-CoV-2 S protein and efficiently supported the pseudotyped virus infection. These findings have important implications for understanding potential natural reservoirs, zoonotic transmission, human-to-animal transmission, and use of animal models.

Importance

SARS-CoV-2 uses human ACE2 as primary receptor for host cell entry. Viral entry mediated by the interaction of ACE2 with spike protein largely determines host range and is the major constraint to interspecies transmission. We examined the receptor activity of 14 ACE2 orthologues and found that wild type and mutant SARS-CoV-2 lacking the furin cleavage site in S protein could utilize ACE2 from a broad range of animal species to enter host cells. These results have important implications in the natural hosts, interspecies transmission, animal models and molecular basis of receptor binding for SARS-CoV-2.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.04.19.048710: (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
    All growth medium was supplemented with 10% fetal bovine serum (FBS), 110 mg/L sodium pyruvate, and 4.5 g/L D-glucose. β-actin antibody and C9 antibody were purchased from Sigma (A2228) and SANTA CRUZ (sc-57432), respectively.
    β-actin
    suggested: (Sigma-Aldrich Cat# A2228, RRID:AB_476697)
    C9
    suggested: None
    A polyclonal antibody against human ACE2 and anti-IDE polyclonal antibody were purchased from R&D Systems (catalog No. AF933 and AF2496, respectively).
    human ACE2
    suggested: None
    anti-IDE
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell lines and antibodies: 293T cells and Lenti-X 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco) (57).
    293T
    suggested: None
    Briefly, HEK293T cells were transfected with plasmid encoding ACE2 with Lipofectamine 2000 (Invitrogen).
    HEK293T
    suggested: None
    Software and Algorithms
    SentencesResources
    Construction of plasmids expressing S, S1 and RBD of SARS-CoV-2: The nucleotide sequence of SARS-CoV-2 S gene was retrieved from NCBI database (isolate Wuhan-Hu-1, GenBank No. MN908947).
    NCBI
    suggested: (NCBI, RRID:SCR_006472)
    Phylogenetic tree was constructed based on the nucleotide sequences of animal ACE2 using the neighbor-joining algorithm implemented in MEGA X.
    MEGA
    suggested: (Mega BLAST, RRID:SCR_011920)
    The models were then aligned and compared to the intact SARS-CoV-2 RBD/ ACE2 complex in PyMOL (The PyMOL Molecular Graphics System, Version 2.0 Schrödinger, LLC).
    PyMOL
    suggested: (PyMOL, RRID:SCR_000305)

    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.

  2. Version published to 10.1101/2020.04.19.048710 on bioRxiv