人間の増強因子TEF-1のクローン化,発現,および転写特性
J H Xiao1, I Davidson, H Matthes
1Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Génie Génétique et de Biologie, Faculté de Médecine, Strasbourg, France.
Cell
|May 17, 1991
まとめ
SV40トランスクリプション強化因子1 (TEF-1) の翻訳は,AUコードンから始まります. クローン化されたTEF-1は特定のDNA要素と結合し,活性を抑制し,細胞特異の制限因子がトランス活性化機能を媒介することを示唆しています.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝子規制 遺伝子規制
- 転写因子 トランスクリプション・ファクター
背景:
- SV40転写増強因子1 (TEF-1) は,遺伝子発現の重要な調節因子である.
- TEF-1の活性,特に細胞特異的調節の正確なメカニズムは,まだ完全に理解されていません.
研究 の 目的:
- TEF-1をコードするcDNAの特徴を特定する.
- TEF-1の翻訳開始部位を調査する.
- 異なる細胞タイプにおけるTEF-1の転写活動と調節のメカニズムを解明する.
主な方法:
- cDNAクローニングとTEF-1のシーケンシング
- in vivoとin vitroのトランスレーションアッセイ.
- SV40エンハンソン (GT-IICとSph) を使用したDNA結合アッセイ.
- HeLaおよびMPC11細胞におけるレポーター遺伝子解析.
- GAL4-TEF-1キメリックタンパク質の構築と試験.
主要な成果:
- TEF-1 cDNAはクローンされ,翻訳はAUUコドンでのみ開始されることが in vivoで示されました.
- クローン化されたTEF-1は,SV40 GT-IICとSph強化機に特異的に結合する.
- リンパ性MPC11細胞では,エンハンソンが不活性であるため,クローン化されたTEF-1は内生TEF-1活動を抑制する.
- 抑制は,GAL4-TEF-1キメラによって示されているように,干渉/抑制によって媒介されます.
- キメリックタンパク質は,HeLa細胞の転写を刺激するが,MPC11細胞の転写を刺激しない.
結論:
- TEF-1の翻訳開始は特定のもので,AUUのコードンで起こります.
- TEF-1のトランス活性化機能は,潜在的に細胞特異性のある,高度に制限される,定位可能な転写中間因子によって媒介される可能性があります.
- 観察された抑制と細胞特異的な活動は,TEF-1機能を制御する複雑な規制メカニズムを強調しています.
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