インフルエンザポリメラーゼとPol II CTDの重要な相互作用の構造的基礎
Maria Lukarska1, Guillaume Fournier2,3,4, Alexander Pflug1
1European Molecular Biology Laboratory, Grenoble Outstation, 71 Avenue des Martyrs, CS 90181, 38042 Grenoble Cedex 9, France.
Nature
|December 22, 2016
まとめ
インフルエンザポリメラーゼは,細胞RNAポリメラーゼIIのC末端領域 (CTD) に,そのPAサブユニットの特定の部位を通して結合する. この相互作用を妨害するとウイルスの転写と複製が妨げられ,抗ウイルス薬の標的となる可能性がある.
科学分野:
- ウイルス学
- 構造生物学
- 分子生物学
背景:
- インフルエンザポリメラーゼ (FluPol) は,ウイルスRNAの転写と複製に不可欠です.
- FluPolは細胞RNAポリメラーゼII (Pol II) と相互作用し,mRNA転写のための5'- cappedプライマーを取得する.
研究 の 目的:
- インフルエンザポリメラーゼとPol II C末端領域 (CTD) の相互作用の構造的基礎を解明する.
- ウイルスの転写と複製におけるこの相互作用の機能的意義を調査する.
主な方法:
- 蝙蝠インフルエンザAのポリメラーゼとPol II CTDペプチドの共結結晶.
- PAサブユニットで特定されたCTD結合残基のサイト指向型変異.
- In vitro RNA合成アッセイと細胞ベースのミニゲノムアッセイ
- リコンビネントインフルエンザウイルスの救出と特徴付け
主要な成果:
- 結晶構造は,多重CTD繰り返しを収容するPAサブユニット内の2つの異なるフォスフォセリン-5 (SeP5) 結合部位を明らかにした.
- SeP5結合残留物の変異はCTD結合を in vitroで弱めたが,細胞ベースの測定ではポリメラーゼ活性を著しく低下させた.
- これらの部位が変異した再結合ウイルスは弱体化し,遺伝的に不安定で,一部の変異は既知の毒性因子と相関していた.
結論:
- インフルエンザポリメラーゼがPol II CTDに直接結合することは,特にSeP5部位において,効率的なウイルス転写に不可欠である.
- インフルエンザポリメラーゼのCTD結合部位は,新しい抗ウイルス療法の開発の潜在的なターゲットです.
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