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For the non\u2010structural proteins (Nsps) of SARS\u2010CoV\u20102, there are limited structural analyses of their naturally occurring mutations. Here, we identified four non\u2010synonymous single\u2010nucleotide polymorphisms (nsSNPs) in the Epsilon lineage of SARS\u2010CoV\u20102 within Nsp15, an endoribonuclease critical for immune evasion. Of these Epsilon nsSNPs, E266Q is in the catalytic domain. This study investigates the effects of this on enzymatic activity, structural stability, and oligomeric assembly by serial crystallography. By solving the structure of the Nsp15 hexamer at room temperature of both Nsp15\u2010E266Q and WT in the P2<jats:sub>1<\/jats:sub> space group to 3\u2009\u00c5, we observed asymmetric motions within its trimer subunits, a feature not visible in previously reported higher\u2010symmetry space groups. These asymmetric motions resemble substrate\u2010induced conformational changes reported in RNA\u2010bound Nsp15 structures, suggesting functional relevance. Biochemical analyses further reveal that Nsp15\u2010E266Q exhibited significantly higher enzymatic activity and thermal stability compared to the wild\u2010type protein. These findings highlight how mutations in Nsp15 contribute to viral replication and immune evasion, offering insights into the molecular mechanisms underlying SARS\u2010CoV\u20102 variant evolution and potential therapeutic strategies.<\/jats:p>","DOI":"10.1002\/pro.70115","type":"journal-article","created":{"date-parts":[[2025,5,24]],"date-time":"2025-05-24T10:50:27Z","timestamp":1748083827000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Functional implications of hexameric dynamics in <scp>SARS<\/scp>\u2010<scp>CoV<\/scp>\u20102 Nsp15"],"prefix":"10.1002","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4665-8831","authenticated-orcid":false,"given":"Manashi","family":"Sonowal","sequence":"first","affiliation":[{"name":"Center for Applied Structural Discovery, Biodesign Institute Arizona State University  Tempe Arizona USA"},{"name":"School of Molecular Sciences Arizona State University  Tempe 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