| 研究課題/領域番号 |
16K08014
|
|---|
| 研究機関 | 北海道大学 |
|---|
研究代表者 |
マンズール ラシッド 北海道大学, 人獣共通感染症リサーチセンター, 特任助教 (90566150)
|
|---|
| 研究期間 (年度) |
2016-04-01 – 2019-03-31
|
| キーワード | Influenza A virus / M2 protein |
|---|
| 研究実績の概要 |
The viruses upon infection hijack the cell machinery and re-route or modulate it to complete their life cycle. Therefore, viruses or their encoded proteins must interact with host proteins for successful completion of their life cycle. Despite the important role of influenza A virus (IAV) M2 protein in virus life cycle such as, vRNP release, virus morphogenesis and budding, very little information on M2-host interactome is available. In this study, we plan to investigate the role of identified M2-interacting host proteins in virus replication.
During this fiscal year, siRNA based screening was conducted using three siRNAs per target gene. Initially screening conditions such as optimal virus dose, siRNA dose, and suitable cell line (293T, A549 etc) were optimized. The host factors were shortlisted by determining the gene knockdown effect on virus infectivity. The effect of gene knockdown on virus replication was considered positive or negative when at least 2/3 siRNAs produced similar effect. So far, knockdown of five genes caused more than thirty percent reduction in infectivity and three genes caused increase in virus infectivity.
|
| 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
Further investigations are being conducted to study the role of shortlisted host proteins in virus replication. Members of solute carrier family proteins i.e. SLC25A5, SLC1A5 are among the shortlisted candidate proteins. At present, there putative role in virus replication is being investigated.
To study the effect of gene knockdown on virus budding, an influenza virus has been generated by reverse genetics. Moreover, a stable cell line expressing M2 protein is being developed to study the effect of gene knockdown on M2 cytotoxicity.
|
| 今後の研究の推進方策 |
1. To continue to investigate the role of shortlisted host proteins in influenza virus replication with relevance to M2 protein function.
2. To optimize 2D electrophoresis for studying the host plasma membrane proteome in M2 expressing cells.
|
| 次年度使用額が生じた理由 |
Since the preparation of rg-virus and stable cell line required for screening of host factors in virus budding were delayed; therefore, the consumables and reagents required could not be purchased during this fiscal year.
|
| 次年度使用額の使用計画 |
The unused amount will be used to continue the study in the next fiscal year. The money will be used for buying the consumables necessary for execution of study such as siRNAs, antibodies, media, plastic ware etc. Additionally, the amount will be used for presenting the research results.
|
