Inhibition of anti-viral stress granule formation by coronavirus endoribonuclease nsp15 ensures efficient virus replication

• Published in: PLoS pathogens 2021-02, Vol.17 (2), p.e1008690-e1008690
• Main Authors: Gao, Bo, Gong, Xiaoqian, Fang, Shouguo, Weng, Wenlian, Wang, Huan, Chu, Hongyan, Sun, Yingjie, Meng, Chunchun, Tan, Lei, Song, Cuiping, Qiu, Xusheng, Liu, Weiwei, Forlenza, Maria, Ding, Chan, Liao, Ying
• Format: Article
• Language: English
• Subjects: Antiviral agents; Biodegradation; Biology and life sciences; Cell Line; Cold shock; Coronaviridae; Coronavirus - immunology; Coronaviruses; COVID-19 - metabolism; COVID-19 - virology; Cytoplasm; Cytoplasmic Granules - immunology; Cytoplasmic Granules - metabolism; Cytoplasmic Granules - virology; Diarrhea; Endoribonucleases - immunology; Endoribonucleases - metabolism; Environmental stress; Epidemics; Feline infectious peritonitis; Gene expression; Heat shock; Humans; Interferon; Interferon-beta - immunology; Interferon-beta - metabolism; Kinases; Medicine and Health Sciences; Middle East respiratory syndrome; Nitric oxide; Oxidative stress; Peritonitis; Perturbation; Polyribosomes; Proteasomes; Puromycin; Replication; Research and Analysis Methods; Ribonucleic acid; RNA; SARS-CoV-2 - metabolism; SARS-CoV-2 - physiology; Severe acute respiratory syndrome; Signal Transduction; Splicing; Transmissible gastroenteritis; Ultraviolet radiation; Viral diseases; Viral Nonstructural Proteins - immunology; Viral Nonstructural Proteins - metabolism; Virus Replication - physiology; Viruses
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/JOURNAL.PPAT.1008690

Cytoplasmic stress granules (SGs) are generally triggered by stress-induced translation arrest for storing mRNAs. Recently, it has been shown that SGs exert anti-viral functions due to their involvement in protein synthesis shut off and recruitment of innate immune signaling intermediates. The largest RNA viruses, coronaviruses, impose great threat to public safety and animal health; however, the significance of SGs in coronavirus infection is largely unknown. Infectious Bronchitis Virus (IBV) is the first identified coronavirus in 1930s and has been prevalent in poultry farm for many years. In this study, we provided evidence that IBV overcomes the host antiviral response by inhibiting SGs formation via the virus-encoded endoribonuclease nsp15. By immunofluorescence analysis, we observed that IBV infection not only did not trigger SGs formation in approximately 80% of the infected cells, but also impaired the formation of SGs triggered by heat shock, sodium arsenite, or NaCl stimuli. We further demonstrated that the intrinsic endoribonuclease activity of nsp15 was responsible for the interference of SGs formation. In fact, nsp15-defective recombinant IBV (rIBV-nsp15-H238A) greatly induced the formation of SGs, along with accumulation of dsRNA and activation of PKR, whereas wild type IBV failed to do so. Consequently, infection with rIBV-nsp15-H238A strongly triggered transcription of IFN-β which in turn greatly affected rIBV-nsp15-H238A replication. Further analysis showed that SGs function as an antiviral hub, as demonstrated by the attenuated IRF3-IFN response and increased production of IBV in SG-defective cells. Additional evidence includes the aggregation of pattern recognition receptors (PRRs) and signaling intermediates to the IBV-induced SGs. Collectively, our data demonstrate that the endoribonuclease nsp15 of IBV interferes with the formation of antiviral hub SGs by regulating the accumulation of viral dsRNA and by antagonizing the activation of PKR, eventually ensuring productive virus replication. We further demonstrated that nsp15s from PEDV, TGEV, SARS-CoV, and SARS-CoV-2 harbor the conserved function to interfere with the formation of chemically-induced SGs. Thus, we speculate that coronaviruses employ similar nsp15-mediated mechanisms to antagonize the host anti-viral SGs formation to ensure efficient virus replication.