Fullerene derivatives with hydrophilic substituents have been shown to exhibit a range of biological activities, including antiviral ones. For a long time, the anti-HIV activity of fullerene derivatives was believed to be due to their binding into the hydrophobic pocket of HIV-1 protease, thereby blocking its activity. Recent work, however, brought new evidence of a novel, protease-independent mechanism of fullerene derivatives' action. We studied in more detail the mechanism of the anti-HIV-1 activity of N,N-dimethyl[70]fulleropyrrolidinium iodide fullerene derivatives. By using a combination of in vitro and cell-based approaches, we showed that these C70 derivatives inhibited neither HIV-1 protease nor HIV-1 maturation. Instead, our data indicate effects of fullerene C70 derivatives on viral genomic RNA packaging and HIV-1 cDNA synthesis during reverse transcription-without impairing reverse transcriptase activity though. Molecularly, this could be explained by a strong binding affinity of these fullerene derivatives to HIV-1 nucleocapsid domain, preventing its proper interaction with viral genomic RNA, thereby blocking reverse transcription and HIV-1 infectivity. Moreover, the fullerene derivatives' oxidative activity and fluorescence quenching, which could be one of the reasons for the inconsistency among reported anti-HIV-1 mechanisms, are discussed herein.

• MeSH
• fullereny metabolismus   farmakologie   MeSH
• genom virový účinky léků   MeSH
• genové produkty gag - virus lidské imunodeficience metabolismus   MeSH
• HEK293 buňky MeSH
• HIV-1 účinky léků   genetika   metabolismus   fyziologie   MeSH
• látky proti HIV metabolismus   farmakologie   MeSH
• lidé MeSH
• nukleokapsida - proteiny metabolismus   MeSH
• reverzní transkripce MeSH
• RNA virová metabolismus   MeSH
• svlékání virového obalu účinky léků   MeSH
• vazba proteinů MeSH
• virion metabolismus   MeSH
• zabalení virového genomu účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

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.

• 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