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| | <StructureSection load='6cya' size='340' side='right'caption='[[6cya]], [[Resolution|resolution]] 2.60Å' scene=''> | | <StructureSection load='6cya' size='340' side='right'caption='[[6cya]], [[Resolution|resolution]] 2.60Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6cya]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/Group_a_rotaviruses Group a rotaviruses]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6CYA OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6CYA FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6cya]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Rotavirus_A Rotavirus A]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6CYA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6CYA FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.6Å</td></tr> |
| - | <tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[6auk|6auk]], [[6cy9|6cy9]]</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">NSP2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=28875 Group A rotaviruses])</td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6cya FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6cya OCA], [https://pdbe.org/6cya PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6cya RCSB], [https://www.ebi.ac.uk/pdbsum/6cya PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6cya ProSAT]</span></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6cya FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6cya OCA], [http://pdbe.org/6cya PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6cya RCSB], [http://www.ebi.ac.uk/pdbsum/6cya PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6cya ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/A2T3N6_9REOV A2T3N6_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] | + | [https://www.uniprot.org/uniprot/NSP2_ROTSH NSP2_ROTSH] 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]<ref>PMID:14699117</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Group a rotaviruses]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Anish, R]] | + | [[Category: Rotavirus A]] |
| - | [[Category: Hu, L]] | + | [[Category: Anish R]] |
| - | [[Category: Prasad, B V.V]] | + | [[Category: Hu L]] |
| - | [[Category: Nsp2]] | + | [[Category: Prasad BVV]] |
| - | [[Category: Rotavirus]]
| + | |
| - | [[Category: S313a mutant]]
| + | |
| - | [[Category: Viral protein]]
| + | |
| Structural highlights
Function
NSP2_ROTSH 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][1]
Publication Abstract from PubMed
The rotavirus (RV) genome is replicated and packaged into virus progeny in cytoplasmic inclusions called viroplasms, which require interactions between RV nonstructural proteins NSP2 and NSP5. How viroplasms form remains unknown. We previously found two forms of NSP2 in RV-infected cells: a cytoplasmically dispersed dNSP2, which interacts with hypophosphorylated NSP5; and a viroplasm-specific vNSP2, which interacts with hyperphosphorylated NSP5. Other studies report that CK1alpha, a ubiquitous cellular kinase, hyperphosphorylates NSP5, but requires NSP2 for reasons that are unclear. Here we show that silencing CK1alpha in cells before RV infection resulted in (i) >90% decrease in RV replication, (ii) disrupted vNSP2 and NSP5 interaction, (iii) dispersion of vNSP2 throughout the cytoplasm, and (iv) reduced vNSP2 protein levels. Together, these data indicate that CK1alpha directly affects NSP2. Accordingly, an in vitro kinase assay showed that CK1alpha phosphorylates serine 313 of NSP2 and triggers NSP2 octamers to form a lattice structure as demonstrated by crystallographic analysis. Additionally, a dual-specificity autokinase activity for NSP2 was identified and confirmed by mass spectrometry. Together, our studies show that phosphorylation of NSP2 involving CK1alpha controls viroplasm assembly. Considering that CK1alpha plays a role in the replication of other RNA viruses, similar phosphorylation-dependent mechanisms may exist for other virus pathogens that require cytoplasmic virus factories for replication.
Phosphorylation cascade regulates the formation and maturation of rotaviral replication factories.,Criglar JM, Anish R, Hu L, Crawford SE, Sankaran B, Prasad BVV, Estes MK Proc Natl Acad Sci U S A. 2018 Dec 3. pii: 1717944115. doi:, 10.1073/pnas.1717944115. PMID:30509975[2]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Carpio RV, González-Nilo FD, Jayaram H, Spencer E, Prasad BV, Patton JT, Taraporewala ZF. Role of the histidine triad-like motif in nucleotide hydrolysis by the rotavirus RNA-packaging protein NSP2. J Biol Chem. 2004 Mar 12;279(11):10624-33. PMID:14699117 doi:10.1074/jbc.M311563200
- ↑ Criglar JM, Anish R, Hu L, Crawford SE, Sankaran B, Prasad BVV, Estes MK. Phosphorylation cascade regulates the formation and maturation of rotaviral replication factories. Proc Natl Acad Sci U S A. 2018 Dec 3. pii: 1717944115. doi:, 10.1073/pnas.1717944115. PMID:30509975 doi:http://dx.doi.org/10.1073/pnas.1717944115
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