A novel mechanism of enhanced transcription activity and fidelity for influenza A viral RNA-dependent RNA polymerase
| العنوان: | A novel mechanism of enhanced transcription activity and fidelity for influenza A viral RNA-dependent RNA polymerase |
|---|---|
| المؤلفون: | Xinzhou Xu, Leo L.M. Poon, Zixi Dai, Lu Zhang, Julie Tung Sem Chu, Peter Pak-Hang Cheung, Xuhui Huang, Tin Hang Chong, Alex Wing Hong Chin, Yuqing Wang |
| المصدر: | Nucleic Acids Research |
| بيانات النشر: | Oxford University Press (OUP), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Transcription, Genetic, AcademicSubjects/SCI00010, viruses, Protein subunit, Mutant, Sequence alignment, Biology, Madin Darby Canine Kidney Cells, Viral Proteins, chemistry.chemical_compound, Dogs, Transcription (biology), Catalytic Domain, RNA polymerase, Genetics, Animals, Nucleic Acid Enzymes, Rational design, High-Throughput Nucleotide Sequencing, RNA, RNA-Dependent RNA Polymerase, Cell biology, Amino Acid Substitution, chemistry, Influenza A virus, Mutation, Nucleoside triphosphate, Sequence Alignment |
| الوصف: | During RNA elongation, the influenza A viral (IAV) RNA-dependent RNA polymerase (RdRp) residues in the active site interact with the triphosphate moiety of nucleoside triphosphate (NTP) for catalysis. The molecular mechanisms by which they control the rate and fidelity of NTP incorporation remain elusive. Here, we demonstrated through enzymology, virology and computational approaches that the R239 and K235 in the PB1 subunit of RdRp are critical to controlling the activity and fidelity of transcription. Contrary to common beliefs that high-fidelity RdRp variants exert a slower incorporation rate, we discovered a first-of-its-kind, single lysine-to-arginine mutation on K235 exhibited enhanced fidelity and activity compared with wild-type. In particular, we employed a single-turnover NTP incorporation assay for the first time on IAV RdRp to show that K235R mutant RdRp possessed a 1.9-fold increase in the transcription activity of the cognate NTP and a 4.6-fold increase in fidelity compared to wild-type. Our all-atom molecular dynamics simulations further elucidated that the higher activity is attributed to the shorter distance between K235R and the triphosphate moiety of NTP compared with wild-type. These results provide novel insights into NTP incorporation and fidelity control mechanisms, which lay the foundation for the rational design of IAV vaccine and antiviral targets. |
| تدمد: | 1362-4962 0305-1048 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::94e6798a231190bd3b6ebbb9336c365b https://doi.org/10.1093/nar/gkab660 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....94e6798a231190bd3b6ebbb9336c365b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13624962 03051048 |
|---|