Targeting of Protein Kinase CK2 Elicits Antiviral Activity on Bovine Coronavirus Infection

Ailyn C Ramón,Yassel Ramos,Viviana Falcón,Silvio E Perea,Arielis Rodríguez-Ulloa,Evelin Caballero,Daylén Aguilar,Yasser Perera,Dania Vázquez-Blomquist,Mauro Rosales,Ke Yang,George V Pérez,María Pilar Rodríguez-Moltó

doi:10.3390/v14030552

Ailyn C Ramón, Yassel Ramos + Show 11 more

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https://doi.org/10.3390/v14030552

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Abstract

Coronaviruses constitute a global threat to the human population; therefore, effective pan-coronavirus antiviral drugs are required to tackle future re-emerging virus outbreaks. Protein kinase CK2 has been suggested as a promising therapeutic target in COVID-19 owing to the in vitro antiviral activity observed after both pharmacologic and genetic inhibition of the enzyme. Here, we explored the putative antiviral effect of the anti-CK2 peptide CIGB-325 on bovine coronavirus (BCoV) infection using different in vitro viral infected cell-based assays. The impact of the peptide on viral mRNA and protein levels was determined by qRT-PCR and Western blot, respectively. Finally, pull-down experiments followed by Western blot and/or mass spectrometry analysis were performed to identify CIGB-325-interacting proteins. We found that CIGB-325 inhibited both the cytopathic effect and the number of plaque-forming units. Accordingly, intracellular viral protein levels were clearly reduced after treatment of BCoV-infected cells, with CIGB-325 determined by immunocytochemistry. Pull-down assay data revealed the physical interaction of CIGB-325 with viral nucleocapsid (N) protein and a group of bona fide CK2 cellular substrates. Our findings evidence in vitro antiviral activity of CIGB-325 against bovine coronavirus as well as some molecular clues that might support such effect. Altogether, data provided here strengthen the rationale of inhibiting CK2 to treat betacoronavirus infections.

Highlights

Coronaviruses (CoVs) comprise a diverse group of enveloped positive-strand RNA viruses in the family Coronaviridae that infect a wide variety of hosts, including mammals and birds, and cause several respiratory and enteric diseases [1,2]
A decade later, a novel lethal zoonotic disease appeared in the Arabian Peninsula, caused by the Middle East respiratory syndrome coronavirus (MERS-CoV), with a 36.1% mortality rate according to the World Health Organization (WHO) [5]
Since previous studies in SARS-CoV-2 infection had shown the antiviral activity of the CK2 inhibitor CX-4945 [12], we tested its putative anti-bovine coronavirus (BCoV) activity

Summary

Introduction

Coronaviruses (CoVs) comprise a diverse group of enveloped positive-strand RNA viruses in the family Coronaviridae that infect a wide variety of hosts, including mammals and birds, and cause several respiratory and enteric diseases [1,2]. These viruses include infectious bronchitis virus (IBV) in chickens, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), swine acute diarrhea syndrome-CoV (SADS-CoV), bovine coronavirus (BCoV), and many others [3]. In 2002, a human pathogenic CoV, severe acute respiratory syndrome (SARS-CoV), caused more than 8000 human infections in 2002–2003, with a case fatality about 10% [4]. A novel coronavirus SARS-CoV-2 emerged from Wuhan, China, in December 2019 and three months later became a pandemic disease with more than 350 million people infected and 5 million deaths across the world (www.worldometers.info/coronavirus, accessed on 31 January 2022)

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