2020 Fiscal Year Final Research Report
Identification of inter-segmental interaction regions of influenza virus genomes using antisense oligonucleotide probes
| Project/Area Number |
18K07149
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Review Section |
Basic Section 49060:Virology-related
|
|---|
| Research Institution | Kitasato University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2018-04-01 – 2021-03-31
|
| Keywords | インフルエンザウイルス / RNAウイルス / 選択的分節集合 / パッケージングシグナル / 機能的核酸 / 核酸プローブ / ウイルスライブラリ / RNP複合体 |
|---|
| Outline of Final Research Achievements |
The genomic RNA of influenza A viruses is segmented, and eight distinct segments are selectively assembled and packaged into a single progeny virion. However, the packaging signal (PS) sequences involved in the selective segment assembly are unclear. This research project aimed to identify these PS sequences and elucidate the molecular mechanisms of selective packaging. Focusing on the 5'-end of the sixth segment, we inferred the PS region based on the inhibition of binding of antisense oligonucleotide probes, and attempted to identify the PS sequence by detecting viral growth inhibition and packaging defects caused by base substitutions. Using three virus libraries in which adjacent regions of 15 bases each were randomly substituted with complementary bases, nine essential bases were finally identified, and a method for analyzing PS sequences at single-nucleotide resolution were established.
|
| Free Research Field |
ウイルス学
|
| Academic Significance and Societal Importance of the Research Achievements |
これまで各分節末端を対象として選択的パッケージングに必要なPS領域の同定が試みられてきた。しかしPS領域はタンパク質コード領域(CDS)と一部重複するため、1塩基単位の解析が困難であった。本課題ではPSと重複するCDSを二重化し機能分離することで、PS領域へ任意の塩基変異を導入可能にした。アンチセンスプローブによる分節間相互作用領域の推定など複数の手法と組み合わせることで、選択的パッケージングに必要な塩基の特定が可能となった。将来、この手法を用いて全分節のPS配列が決定できれば、分節交雑ゲノムの安定性を塩基配列から予測し、新型ウイルス発生の可能性を見積もることが可能となるかもしれない。
|
