Interferons and viruses induce a novel primate-specific isoform dACE2 and not the SARS-CoV-2 receptor ACE2
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Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes COVID-19, utilizes angiotensin-converting enzyme 2 (ACE2) for entry into target cells. ACE2 has been proposed as an interferon-stimulated gene (ISG). Thus, interferon-induced variability in ACE2 expression levels could be important for susceptibility to COVID-19 or its outcomes. Here, we report the discovery of a novel, primate-specific isoform of ACE2 , which we designate as deltaACE2 (dACE2) . We demonstrate that dACE2 , but not ACE2 , is an ISG. In vitro , dACE2, which lacks 356 N-terminal amino acids, was non-functional in binding the SARS-CoV-2 spike protein and as a carboxypeptidase. Our results reconcile current knowledge on ACE2 expression and suggest that the ISG-type induction of dACE2 in IFN-high conditions created by treatments, inflammatory tumor microenvironment, or viral co-infections is unlikely to affect the cellular entry of SARS-CoV-2 and promote infection.
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SciScore for 10.1101/2020.07.19.210955: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
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Antibodies Sentences Resources Fixed cells were incubated with rabbit anti-FLAG antibody (1:250 dilution, Thermo Fisher) overnight, washed and then stained with anti-rabbit Alexa Fluor 680 (1:500 dilution, Thermo Fisher). anti-FLAGsuggested: Noneanti-rabbitsuggested: NoneCell lysates were analyzed by Western blots with C-terminal anti-ACE2 antibody (Abcam), and the amounts of recombinant ACE2-GFP and dACE2-GFP proteins in lysates were quantified by densitometry analysis (Imagelab, BD Biosciences) of Western blots. anti-ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources Cells listed in Table S3 were infected in … SciScore for 10.1101/2020.07.19.210955: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
NIH rigor criteria are not applicable to paper type.Table 2: Resources
Antibodies Sentences Resources Fixed cells were incubated with rabbit anti-FLAG antibody (1:250 dilution, Thermo Fisher) overnight, washed and then stained with anti-rabbit Alexa Fluor 680 (1:500 dilution, Thermo Fisher). anti-FLAGsuggested: Noneanti-rabbitsuggested: NoneCell lysates were analyzed by Western blots with C-terminal anti-ACE2 antibody (Abcam), and the amounts of recombinant ACE2-GFP and dACE2-GFP proteins in lysates were quantified by densitometry analysis (Imagelab, BD Biosciences) of Western blots. anti-ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources Cells listed in Table S3 were infected in duplicates or triplicates with SeV (7.5×105 CEID50/ml) for 12 or 24 hrs as previously described19–21. Infections with SARS-CoV-2 in colon cancer cell lines (Caco2 and T84) and lung cancer cell line (Calu3) and colon and ileum organoid cultures were previously described23. Caco2suggested: NoneOrganoids were infected with 3×105 FFU of the virus, based on titers in Vero E6 cells. Vero E6suggested: RRID:CVCL_XD71)T84 and Caco-2 cells were treated with IFN-γ (2 ng/ml) for 48 hrs, followed by cell harvesting, RNA extraction and expression analysis. Caco-2suggested: NoneSoftware and Algorithms Sentences Resources Cell lines were either used within 6 months after purchase or were periodically authenticated by microsatellite fingerprinting (AmpFLSTR Identifiler, Thermo Fisher) by the Cancer Genomics Research Laboratory/NCI). Cancer Genomics Research Laboratory/NCIsuggested: NoneRNA-seq analysis of data from NCBI Sequence Read Archive (SRA) and TCGA: RNA-seq datasets (Table S6) were downloaded from NCBI SRA using SRA tools. NCBI Sequence Read Archivesuggested: (NCBI Sequence Read Archive (SRA, RRID:SCR_004891)The FASTQ files were compressed using GZIP and aligned with STAR version 7.1.3a to the GRChg38/hg38 genome assembly. GZIPsuggested: (Gzip, RRID:SCR_009291)STARsuggested: (STAR, RRID:SCR_015899)BAM files were indexed and sliced using SAM tools to include 51.6 Kb of the ACE2 genomic region (chrX:15,556,393-15,608,016, hg38). SAMsuggested: (SAM, RRID:SCR_010951)https://www.cbioportal.org/). https://www.cbioportal.org/suggested: (cBioPortal, RRID:SCR_014555)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.
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