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In vivo and in vitro Studies of Adaptor-clathrin Interaction
Published on: January 27, 2011
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70kdのアデニリルサイクラゼ関連タンパク質をコードするS. cerevisiaeの遺伝子であるCAPのクローニングと特徴付け
J Field1, A Vojtek, R Ballester
1Cold Spring Harbor Laboratory, New York 11724.
Cell
|April 20, 1990
まとめ
サイクラゼ関連タンパク質 (CAP) は,酵母におけるRAS反応性アデニリルサイクラゼシグナル伝達に不可欠である. CAPが欠けているミュータントは,成長欠陥と形態学的変化も表しており,より広範な細胞役割を示唆しています.
科学分野:
- 分子生物学は分子生物学である.
- 細胞生物学 細胞生物学
- バイオケミストリー バイオケミストリー
背景:
- Saccharomyces cerevisiaeのアデニリルサイクラゼは,複数のサブユニットを持つ酵素である.
- RAS/アデニリルサイクラゼ経路は,様々な刺激に対する細胞の反応を調節する.
研究 の 目的:
- 酵母アデニリルサイクラゼのサイクラゼ関連タンパク質 (CAP) のサブユニットをクローンし,特徴づけること.
- RAS媒介シグナル伝達および他の細胞プロセスにおけるCAPの役割を調査する.
主な方法:
- E. coli の酵母発現ライブラリを使用して,CAP cDNA のクローニング.
- 浄化されたCAPに対する抗菌剤によるスクリーニング.
- CAP欠乏酵母膜におけるアデニリルサイクラースの活性に関する分析.
- CAPミュータントのフェノタイプ分析.
主要な成果:
- 526アミノ酸のCAPタンパク質をコードするcDNAが分離され,既知の配列との同質性は示されなかった.
- CAPが欠けていた酵母は,RAS2刺激によるアデニリルサイクラースの活性が低下したことを示した.
- CAP欠乏変異体は,温度感受性,豊かな環境で成長できない,窒素飢餓感受性,細胞形態の変化を示した.
結論:
- CAPは,酵母におけるRAS反応性アデニリルサイクラゼシグナル伝達経路の重要な構成要素である.
- CAPはRASシグナル伝達を超えて,成長,形態,ストレス反応に影響を与える役割を果たします.
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