We describe the expression and purification of an active recombinant Zika virus RNA-dependent RNA polymerase (RdRp). Next, we present the development and optimization of an in vitro assay to measure its activity. We then applied the assay to selected triphosphate analogs and discovered that 2'-C-methylated nucleosides exhibit strong inhibitory activity. Surprisingly, also carbocyclic derivatives with the carbohydrate locked in a North-like conformation as well as a ribonucleotide with a South conformation exhibited strong activity. Our results suggest that the traditional 2'-C-methylated nucleosides pursued in the race for anti-HCV treatment can be superseded by brand new scaffolds in the case of the Zika virus.
- MeSH
- adenosintrifosfát analogy a deriváty chemie MeSH
- antivirové látky farmakologie MeSH
- inhibitory enzymů farmakologie MeSH
- lidé MeSH
- molekulární konformace MeSH
- nukleosidy chemie farmakologie MeSH
- objevování léků MeSH
- RNA-dependentní RNA-polymerasa antagonisté a inhibitory genetika izolace a purifikace MeSH
- virus zika účinky léků enzymologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Zika virus is considered a major global threat to human kind. Here, we present a crystal structure of one of its essential enzymes, the methyltransferase, with the inhibitor sinefungin. This structure, together with previously solved structures with bound substrates, will provide the information needed for rational inhibitor design. Based on the structural data we suggest the modification of the adenine moiety of sinefungin to increase selectivity and to covalently link it to a GTP analogue, to increase the affinity of the synthesized compounds.
We report the development of a novel method for detection of Bartonella DNA in ixodid ticks. The assay is based on a specific amplification of a part of 16S rRNA gene and 16S-23S rRNA intergenic spacer region of Bartonella sp. by nested PCR and Southern blot hybridization with specific DNA probe; the method is highly sensitive and specific. The screening of 327 unfed ticks collected in different urban and suburban areas of Czechia in 2003-2005 revealed the presence of Bartonella DNA in four Ixodes ricinus individuals (1.2%), two males, one female and one nymph.
- MeSH
- Bartonella genetika izolace a purifikace MeSH
- DNA bakterií genetika izolace a purifikace MeSH
- ELISA metody využití MeSH
- finanční podpora výzkumu jako téma MeSH
- klíšťata mikrobiologie MeSH
- lidé MeSH
- polymerázová řetězová reakce metody využití MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
