2017 Fiscal Year Final Research Report
Anti-Inflienza drug screening targetted by cap-snatching mechanism
| Project/Area Number |
15K08502
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Virology
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| Research Institution | Kitasato University |
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Principal Investigator |
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Keywords | cap structure / antiinfluenza drug / cap-snatching |
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| Outline of Final Research Achievements |
This cap-dependent endonuclease activity termed as cap snatching may provide a unique target for novel anti-influenza viral agents. For the screening of candidate inhibitors for cap-snatching activity, it is essential to establish a method to efficiently produce Cap1-RNA substrate and a convenient assay system for cap-snatching activity. A short 3’-biotinylated RNA oligonucleotide was prepared by an in vitro transcription system utilizing T7 RNA polymerase. Purified 3’-biotinylated RNA oligonucleotide by C18 cartridge column was subjected to a sequential capping reaction with recombinant enzymes of vaccinia virus D1R containing with RNA 5’-triphosphatase and mRNA guanylyltransferase, yeast
ABD1 as (guanine-N7)-methyltransferase, and VP39 as (nucleoside-2'-O-)-methyltransferase. Cap-snatching activity could be analyzed a pull-down assay based on the streptavidin-biotin interaction, yielding results consistent with those obtained by polyacrylamide gel electrophoresis.
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| Free Research Field |
ウイルス学関連
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