タンパク質因子であるASFは,SV40初期プレ-mRNAの細胞特異的な代替スプライシングをインビトロで制御する
1Department of Biological Sciences, Columbia University, New York, New York 10027.
Cell
|July 13, 1990
まとめ
293の細胞内の特定のタンパク質因子は,シミアンウイルス40 (SV40) のスプライシング中に小さなt mRNAの生成を高めます. この要因は選択的に他の哺乳類の細胞と異なる代替スプライシングを促進します.
科学分野:
- 分子生物学は分子生物学である.
- RNA スプライシング
- ウイルス学 ウイルス学 ウイルス学
背景:
- シミアンウイルス40 (SV40) の初期のプレ-mRNAは,大きなT型と小さなt型mRNAを生成するために,代替スプライシングを受けます.
- ヒト胚性腎臓293細胞では,他の哺乳類の細胞と比較して,小さなtと大きなTのmRNAの比率が高いことが観察されています.
研究 の 目的:
- 293個の細胞におけるSV40初期プレ-mRNAの異なるスプライシング比率の分子基礎を調査する.
- 潜在的な細胞特異のスプライシング因子を特定し,特徴づけること.
主な方法:
- 293およびHeLa細胞の核抽出物を用いたインビトロスプライシングアッセイ.
- splicing factorを分離し,浄化するために,in vitro補完測定法を行います.
- 精製された因子の生化学的特徴 (熱感性,核酸抵抗性).
主要な成果:
- 293個の細胞からの核抽出物は,HeLa細胞抽出物よりも,著しく高いsmall t splicing活性を示した.
- 熱に敏感でヌクレアゼに抵抗する因子を,選択的に小型のt mRNAの産生を増加させる293の細胞から浄化した.
- アクティブ・スプライシング・ファクターは,30~35kdのタンパク質であるようです.
結論:
- 細胞特異タンパク質因子A293は,SV40の代替プレ-mRNAスプライシングを調節する.
- この因子は,大TmRNAよりも小TmRNAの生成を優先的に促進する.
- 特定された因子は,特定の細胞文脈における代替スプライシングの重要な調節因子である.
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