Rotavirus genomes are distributed between 11 distinct RNA molecules, all of which must be selectively copackaged during virus assembly. This likely occurs through sequence-specific RNA interactions facilitated by the RNA chaperone NSP2. Here, we report that NSP2 autoregulates its chaperone activity through its C-terminal region (CTR) that promotes RNA-RNA interactions by limiting its helix-unwinding activity. Unexpectedly, structural proteomics data revealed that the CTR does not directly interact with RNA, while accelerating RNA release from NSP2. Cryo-electron microscopy reconstructions of an NSP2-RNA complex reveal a highly conserved acidic patch on the CTR, which is poised toward the bound RNA. Virus replication was abrogated by charge-disrupting mutations within the acidic patch but completely restored by charge-preserving mutations. Mechanistic similarities between NSP2 and the unrelated bacterial RNA chaperone Hfq suggest that accelerating RNA dissociation while promoting intermolecular RNA interactions may be a widespread strategy of RNA chaperone recycling.

• Klíčová slova
• RNA chaperones, genome assembly, ribonucleoproteins, rotavirus,
• MeSH
• elektronová kryomikroskopie MeSH
• genom virový genetika   MeSH
• molekulární chaperony metabolismus   MeSH
• molekulární modely MeSH
• proteiny vázající RNA metabolismus   MeSH
• ribonukleoproteiny metabolismus   MeSH
• RNA virová genetika   MeSH
• Rotavirus genetika   růst a vývoj   metabolismus   MeSH
• sbalování RNA genetika   MeSH
• virové nestrukturální proteiny metabolismus   MeSH
• zabalení virového genomu genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• molekulární chaperony MeSH
• proteiny vázající RNA MeSH
• ribonukleoproteiny MeSH
• RNA virová MeSH
• virové nestrukturální proteiny MeSH

Human rotaviruses present in faecal specimens of patients with gastroenteritis were isolated and adapted to growth in MA-104 cells in roller tubes using trypsin pretreatment. Storage temperature-dependent differences were found in the extent of cytopathic effect. Multiplication of viruses in cell cultures was confirmed by immunoelectron microscopy and ELISA. Electrophoresis of rotavirus RNA in polyacrylamide gel was used to verify the identity of the electropherotypes determined in original faecal specimens and in final cell passages.

• MeSH
• buněčné linie MeSH
• cytopatogenní efekt virový MeSH
• feces mikrobiologie   MeSH
• gastroenteritida mikrobiologie   MeSH
• Haplorrhini MeSH
• kultivace virů MeSH
• lidé MeSH
• RNA virová analýza   MeSH
• rotavirové infekce diagnóza   mikrobiologie   MeSH
• Rotavirus růst a vývoj   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Slovenská republika MeSH
• Názvy látek
• RNA virová MeSH

Twenty rotavirus strains were isolated in 1991-92 from 60 faecal samples collected from diarrhoeic piglets in the Czech and Slovak Republics. Three isolates were adapted to the growth in the cell line MA-104 and produced cytopathic effect. Rotavirus was demonstrated by immunofluorescence test, electron microscopy, polyacrylamide gel electrophoresis, immunoperoxidase test and ELISA.

• MeSH
• buněčné linie MeSH
• cytopatogenní efekt virový MeSH
• feces mikrobiologie   MeSH
• nemoci prasat mikrobiologie   MeSH
• prasata MeSH
• průjem mikrobiologie   veterinární   MeSH
• RNA virová analýza   MeSH
• rotavirové infekce mikrobiologie   veterinární   MeSH
• Rotavirus genetika   růst a vývoj   izolace a purifikace   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Slovenská republika MeSH
• Názvy látek
• RNA virová MeSH