9oft
Rotavirus NSP2 WT
Structural highlights
FunctionA2T3N6_9REOV Participates in replication and packaging of the viral genome. Plays a crucial role, together with NSP5, in the formation of virus factories (viroplasms) which are large inclusions in the host cytoplasm where replication intermediates are assembled and viral RNA replication takes place. Displays ssRNA binding, NTPase, RNA triphosphatase (RTPase) and ATP-independent helix-unwinding activities. The unwinding activity may prepare and organize plus-strand RNAs for packaging and replication by removing interfering secondary structures. The RTPase activity plays a role in the removal of the gamma-phosphate from the rotavirus RNA minus strands of dsRNA genome segments.[HAMAP-Rule:MF_04089][RuleBase:RU364009] Publication Abstract from PubMedRotaviruses (RVs) induce the formation of cytoplasmic viral factories, termed viroplasms, which are the sites of early particle assembly and viral RNA synthesis. The RV octameric nonstructural protein 2 (NSP2) plays critical, albeit incompletely understood, roles during viroplasm biogenesis. Previous work by our lab demonstrated that a RV bearing a lysine-to-glutamic acid (K294E) change in the flexible C-terminus of NSP2 exhibits defects in viral replication and induces smaller, more numerous viroplasms as compared to the wildtype (WT) virus. In this study, we sought to better understand if/how this K294E amino acid change altered the structure and/or dynamics of the NSP2 protein. We first determined the X-ray crystal structures of untagged, recombinant NSP2(K294E) and NSP2(WT). We found that both proteins formed highly similar octamers and crystallized in the I422 space group. To better understand the possible impacts of the K294E change on the conformations and backbone flexibility of NSP2, we performed molecular dynamics simulations. The results showed that NSP2(K294E) adopted distinct C-terminal conformations relative to NSP2(WT) and had subtle flexibility differences. Most notably, the data suggest that the K294E change stabilized a rare C-terminal conformation that was only infrequently sampled by NSP2(WT). This shift in conformational preference may help explain why NSP2(K294E) displayed decreased capacity to mediate robust viroplasm formation during RV infection. These results provide mechanistic insights into how a single amino acid change in the NSP2 C-terminus can have large effects on structural ensemble, shedding light on features of the protein that underpin RV viroplasm formation. K294E change in the rotavirus factory forming protein NSP2 stabilizes a rare C-terminal conformation.,Nichols SL, Hollis T, Salsbury FR, Esstman SM J Biomol Struct Dyn. 2025 Sep 26:1-17. doi: 10.1080/07391102.2025.2563689. PMID:40999894[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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