| 研究実績の概要 |
Though primarily an ion channel, the M2 protein is not only involved in various steps of viral replication but also affects the host cellular functions in ion channel activity-dependent and -independent manners. While en route to the cell membrane, the M2-ectodomain and cytoplasmic tail are exposed to vesicular/organellar and cytoplasmic environments, where they may interact with various cellular proteins. Therefore, in this study, we plan to investigate the role of M2-interacting host proteins in virus replication.
The M2 protein functions at various steps of virus replication, therefore, the roles of identified host proteins were further evaluated by comparing the infectivity rates of cells and virus release by plaque assay. For this purpose, a plaque assay suitable for large scale screening was developed. Furthermore, to assess the role of host factors in M2-mediated functions, stable cell lines expressing M2 protein were generated using 239 and A549 cell lines and characterized. An assay to assess the M2-ion channel function was also developed and optimized. Moreover, optimal technique for plasma membrane proteome from mock and M2-expressing cells was developed and attempts were made to analyze by 2D-electrophortesis.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The role of identified host proteins in influenza A virus M2 intracellular transport to plasma membrane is being assessed using M2 expressing stable cell lines. Alongside, further investigations are also being conducted to study the role of shortlisted host proteins in M2-function related stages of virus replication.
Presently, 2D-electrophoresis for studying membrane proteome is being optimized.
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| 次年度使用額が生じた理由 |
The manuscript submission was planned, but got shifted to next fiscal year. The M2 protein is cytotoxic, so development of M2-expressing stable cell lines took time, therefore, the consumables and reagents required could not be purchased during this fiscal year.
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