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High-throughput Purification of Affinity-tagged Recombinant Proteins
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機能的プリイニシテーション複合体の組み立てにおけるトランスクリプション因子ATFの役割に関する分析
T W Hai1, M Horikoshi, R G Roeder
1Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Cell
|September 23, 1988
まとめ
転写因子ATFは,機能的プレイニシアーション複合体の形成に一時的な役割を果たします. ATFおよび一般的な転写因子を含むこれらの複合体の早期組み立てにより,ATF封鎖に耐性があります.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝子規制 遺伝子規制
- バイオケミストリー バイオケミストリー
背景:
- 添付論文では,DNAase I足跡を用いて,活性化トランスクリプション因子 (ATF) と一般的なトランスクリプション開始因子の相互作用を分析した.
- 転写の開始におけるATFの正確な役割を理解することは,遺伝子調節機構の解読に不可欠です.
研究 の 目的:
- トランスクリプションアッセイを用いて,機能的なプレイニシアーション複合体の組み立てにおけるATFの役割を調査する.
- ATFがプレイニシテーション複合体の形成に参加する段階とその他の転写因子への依存度を決定する.
主な方法:
- トランスクリプションアッセイは,E4遺伝子発現をモニタリングするために使用されました.
- ATF結合部位を含むオリゴヌクレオチドは,ATFを隔離し,転写への影響を評価するために使用されました.
- 複合体の形成を研究するために,核抽出物によるE4プロモーターの予備化が行われました.
主要な成果:
- ATF結合部位オリゴヌクレオチドの添加は,ATFを隔離することによってE4転写を阻害しました.
- E4プロモーターのプレインキュベーションにより,ATFオリゴヌクレオチドによる阻害に転写が耐性となり,早期複合体の形成を示唆した.
- このオリゴヌクレオチド耐火性複合体の形成は,ATF,RNAポリメラーゼII,TFIIB,TFIIDに依存しており,協力的な相互作用を示唆しています.
- TFIIEは,転写活動には必要だったが,耐火性の複合体の組み立てには必要ではなかった.
結論:
- ATFは,プレイニシアチブ・コンプレックス・アセンブリの初期段階に関与しています.
- 機能的プレイニシアチブ複合体の形成には,複数の転写因子間の協力的な相互作用が含まれます.
- ATFは,機能的なプリイニシテーション複合体の組み立てのために一時的にのみ必要であり,転写開始におけるダイナミックな役割を強調する可能性があります.
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