ANNOUNCEMENTS
The SARS-CoV-2 Nucleocapsid protein (N protein) plays an essential role in viral genome packaging and pathogenesis. The N protein comprises three intrinsically disordered regions (IDR 1-3), an RNA binding domain, and a C-terminal dimerization domain. Among these, the N-terminal RNA-binding domain (RBD) of SARS-CoV-2 Nucleocapsid protein is crucial for RNA binding and is heavily phosphorylated. To understand the role of phosphorylation in the RNA binding, we initiated in-silico analysis and subsequently, the production of the RNA binding domain (RBD) of N protein. In this study, we characterized the phosphorylation sites present in the RBD through in-silico analysis. We also identified the kinases that phosphorylate N protein. Our studies show that the phosphorylation sites are mostly localized to the RBD and Intrinsically Disordered Region 2 of N protein. Hence, to evaluate the role of phosphorylation of the RBD protein, we amplified and cloned RBD of N protein into a pETRP1 vector to overexpress in the E. coli.
Keywords: N protein, RNA-Binding Domain, Phosphorylation, Cloning, Expression.
