SARS-CoV-2 nucleocapsid protein dually regulates innate immune responses
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Abstract
The recently emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the ongoing global pandemic of COVID-19, may trigger immunosuppression in the early stage and a cytokine storm in the late stage of infection, however, the underlying mechanisms are not well understood. Here we demonstrated that the SARS-CoV-2 nucleocapsid (N) protein dually regulated innate immune responses, i.e., the low-dose N protein suppressed type I interferon (IFN-I) signaling and inflammatory cytokines, whereas high-dose N protein promoted IFN-I signaling and inflammatory cytokines. Mechanistically, the SARS-CoV-2 N protein interacted with the tripartite motif protein 25 (TRIM25), thereby dually regulating the phosphorylation and nuclear translocation of IRF3, STAT1 and STAT2. Additionally, low-dose N protein combined with TRIM25 could suppress retinoic acid-inducible gene I (RIG-I) ubiquitination and activation. Our findings revealed a regulatory mechanism of innate immune responses by the SARS-CoV-2 N protein, which would contribute to understanding the pathogenesis of SARS-CoV-2 and other SARS-like coronaviruses, and development of more effective strategies for controlling COVID-19.
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SciScore for 10.1101/2021.02.17.431755: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies against STAT1, p-STAT1, and STAT2 were purchased from Abcam (Cambridge, MA, USA); anti-IRF3, anti-HA, anti-GST, anti-Flag, anti-GAPDH, CoraLite 594-conjugated IgG and CoraLite 488-conjugated IgG secondary antibodies were obtained from Proteintech (Rosemont, IL, USA); anti-TRIM25 and anti-pIRF3 antibodies were from Cell Signaling technology (Danvers, MA, USA); anti-pSTAT2 antibody was from Sigma-Aldrich (St. Louis, MO, USA). STAT1suggested: Nonep-…SciScore for 10.1101/2021.02.17.431755: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies against STAT1, p-STAT1, and STAT2 were purchased from Abcam (Cambridge, MA, USA); anti-IRF3, anti-HA, anti-GST, anti-Flag, anti-GAPDH, CoraLite 594-conjugated IgG and CoraLite 488-conjugated IgG secondary antibodies were obtained from Proteintech (Rosemont, IL, USA); anti-TRIM25 and anti-pIRF3 antibodies were from Cell Signaling technology (Danvers, MA, USA); anti-pSTAT2 antibody was from Sigma-Aldrich (St. Louis, MO, USA). STAT1suggested: Nonep-STAT1suggested: NoneSTAT2suggested: Noneanti-IRF3suggested: Noneanti-HAsuggested: Noneanti-GSTsuggested: Noneanti-Flagsuggested: Noneanti-GAPDHsuggested: NoneUSA)suggested: Noneanti-TRIM25suggested: Noneanti-pIRF3suggested: Noneanti-pSTAT2suggested: NoneExperimental Models: Cell Lines Sentences Resources Cells and antibodies: Human embryonic kidney cell HEK293T and hepatic carcinoma cell HepG2 were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (HyClone, Logan, UT) containing 10% inactivated fetal bovine serum (BBI, Shanghai, China), penicillin (100 IU/ml) and streptomycin (100 mg/ml) at 37 °C in a 5% CO2 atmosphere. HepG2suggested: NoneBriefly, HEK293T, HepG2 or sg25 cells were transfected with a control plasmid or protein expression plasmids together with the luciferase reporter plasmids using Viafect TM Transfection Reagent (Promega, Madison, WI, USA) or Lipofectamine TM 2000 (Invitrogen, San Diego, CA, USA). HEK293Tsuggested: NoneSoftware and Algorithms Sentences Resources After cells were washed with PBST, they were blocked in 1% BSA and stained with primary antibodies, followed by staining with CoraLite 594- or CoraLite 488-conjugated IgG secondary antibodies (41). CoraLitesuggested: NoneResults 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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 8 and 15. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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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