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*Corresponding Author: Ashraf Albrakati,
SARS-CoV-2 Non-Structural Protein 6 (NSP6) is pivotal for viral replication, but a comprehensive understanding of its evolutionary stability, functional sites and dynamic behavior has been limited. Leveraging a newly established high-confidence 3D model, this study provides an integrative analysis of NSP6’s biology and biophysics.
The validated NSP6 structure was analyzed for conserved regions and evolutionary history using BioEdit and MEGA11 for phylogenetic tree construction. Functional motifs and post-translational modification (PTM) sites were predicted using PROSITE, SMART, MotifFinder and MotifScan. Structural classification was performed using CATH, SCOP and SUPERFAMILY. A 100 ns molecular dynamics (MD) simulation was conducted using GROMACS with the CHARMM27 force field to evaluate structural stability through RMSD, RMSF, Rg and SASA.
Phylogenetic analysis revealed NSP6’s close relationship to bat coronaviruses and identified a single, fully conserved domain across its entire 290-amino-acid length. Motif analysis identified the definitive Coronavirus replicase NSP6 domain and predicted critical PTM sites, including Casein Kinase II phosphorylation and N-myristoylation sites. Structural classification revealed an unexpected homology to cobalamin adenosyltransferase-like folds. The 100 ns MD simulation demonstrated outstanding model stability, with low RMSD (0.2-0.35 nm after 20 ns), a consistent radius of gyration (2.04±0.01 nm) and stable solvent-accessible surface area.
Functional motifs, Molecular dynamics simulation, NSP6, Phylogenetic tree, Post-translational modifications, RMSD, RMSF, SARS-CoV-2