2016 Fiscal Year Annual Research Report
原子間力顕微鏡によるＣ型肝炎ウイルス増殖機構の分子基盤の解明
| Project/Area Number |
16F16390
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| Research Institution | Kyoto University |
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Principal Investigator |
野田 岳志 京都大学, ウイルス・再生医科学研究所, 教授 (00422410)
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| Co-Investigator(Kenkyū-buntansha) |
GILMORE JAMIE 京都大学, ウイルス・再生医科学研究所, 外国人特別研究員
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| Project Period (FY) |
2016-11-07 – 2019-03-31
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| Keywords | Hepatitis C virus / Atomic force microscopy |
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| Outline of Annual Research Achievements |
My recent work has focused on development of high-throughput procedures to extract structural information from RNA molecules imaged with AFM. In addition to HCV, these procedures were applied to other RNA molecules. Findings correlating the structures of 28S rRNA molecules in our AFM images to reported the cryo-EM structure have been submitted for publication. Also, a book chapter discussing the methods for imaging RNA with AFM which was published in February. I have also applied these procedures to influenza virus mRNA molecules, identifying large structures encompassing the splice sites. I have also begun using RNA prediction software in combination with my method to model the secondary structure of these molecules, and I have been able to get high-speed AFM images of RNA dynamics.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Currently, the advancements I have made to my algorithms towards making this a high-throughput approach will facilitate the analysis of the full length HCV genome, and the high speed experiments will facilitate protein-interaction experiments in the future. Also, I am trying to make additional adjustments to the hsAFM sample preparation methods to increase the resolution of the images. The progress on the HCV project has been somewhat impeded by the side projects on influenza and ribosomal RNA, but these studies have helped to provide validation for the method and establish this as a revolutionary new technique for the characterization of RNA structure for a variety of systems.
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| Strategy for Future Research Activity |
My goal for the near future is to continue using the RNA structure web server to refine the structural domains of RNA molecules. So far, we have predicted models for the secondary structure, but it is desirable to model the tertiary structure as well. Further validation of these structure could be obtained through chemical mapping. Also, we have also observed some molecular domains existing in varying conformations. The addition of proteins to the RNA may help to identify if these conformers might affect their ability of these domains to form important biological complexes. The high speed AFM images I have obtained are promising, but we are still working to modulate the mobility of the RNA molecules to obtain optimal resolution.
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