The homology analysis of ACE2 gene and its distinct expression in laboratory and wild animals

  1. Gang Wang
  2. A-Mei Zhang
  3. Binghui Wang
  4. Jianhua Yin
  5. Yue Feng
  6. zulqarnain Baloch
  7. Xueshan Xia

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Abstract

Angiotensin-converting enzyme-2 (ACE2) has been recognized as an entry receptor of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into the host cells while bats has been suspected as natural host of SARS-CoV-2. However, the detail of intermediate host or the route of transmission of SARS-CoV-2 is still unclear. In this study, we analyze the conservation of ACE2 gene in 11 laboratory and wild animals that live in close proximity either with Bats or human and further investigated its RNA and protein expression pattern in wild bats, mice and tree shrew. We verified that the wild-bats and mice were belonged to Hipposideros pomona and Rattus norvegicus, respectively. ACE2 gene is highly conserved among all 11 animals species at the DNA level. Phylogenetic analysis based on the ACE2 nucleotide sequences revealed that wild bat and Tree shrew were forming a cluster close to human. We further report that ACE2 RNA expression pattern is highly species-specific in different tissues of different animals. Most notably, we found that the expression pattern of ACE2 RNA and protein are very different in each animal species. In summary, our results suggested that ACE2 gene is highly conserved among all 11 animals species. However, different relative expression pattern of ACE2 RNA and protein in each animal species is interesting. Further research is needed to clarify the possible connection between different relative expression pattern of ACE2 RNA and protein in different laboratory and wild animal species and the susceptibility to SARS-CoV-2 infection.

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  1. ScreenIT

    SciScore for 10.1101/2021.04.08.439088: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: Ethical statement: In this study, all the animal related to experiments were strictly accomplished following the instruction of Chinese Regulations of Laboratory Animals (Ministry of Science and Technology of the People’s Republic of China) and Laboratory Animal—Requirements of Environment and Housing Facilities (GB 14925-2010, National Laboratory Animal Standardization Technical Committee).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    ACE2 protein primary antibody (BOSTER, Lot#BST19874624) and β-actin protein primary antibody (ABclonal, Lot#9100026001) respectively, and incubated overnight at 4 °C.
    Lot#BST19874624
    suggested: None
    β-actin protein
    suggested: (ImmunoWay Cat# YM3028, RRID:AB_2629465)
    Experimental Models: Organisms/Strains
    SentencesResources
    Oryctolagus cuniculus f. domesticus, Fancy Rat, Golden hamster, Sprague-Dawley Rat and Phodopus sungorus were purchased from local pet animal market.
    Sprague-Dawley
    suggested: RRID:RGD_70508)
    Software and Algorithms
    SentencesResources
    After the sequence was obtained, confirmed to be β-globin gene sequence by BLST, in NCBI database.
    NCBI
    suggested: (NCBI, RRID:SCR_006472)
    Multi-sequence alignment of the ACE2 of the 11 species was performed using ClustalW, the gene sequence comparison and Neighbor-Joining tree were performed with MEGA 7.0 software [15].
    ClustalW
    suggested: (ClustalW, RRID:SCR_017277)
    MEGA
    suggested: (Mega BLAST, RRID:SCR_011920)
    The translation of ACE2 gene sequences of 11 species into amino acid sequences was also completed by MEGA7.0 software, and then we compared the differences in the key sites of the binding of ACE2 protein and SARS-CoV-2Spike protein in these 11 species, which were reported by Fang S et al[15].
    MEGA7.0
    suggested: None
    The conservation of ACE2 key site residues is accomplished by WebLogo[16].
    WebLogo[16
    suggested: None

    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 10.1101/2021.04.08.439088 on bioRxiv