関連する実験動画
Updated: Jun 25, 2026
10:37
Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
Published on: May 10, 2018
FIP1遺伝子は,酵母前mRNAポリアデニレーション因子の成分をコードし,ポリア) ポリメラーゼと直接相互作用する
P J Preker1, J Lingner, L Minvielle-Sebastia
1Department of Cell Biology, University of Basel, Switzerland.
Cell
|May 5, 1995
まとめ
研究者らは,ポリアデニレーションに不可欠な遺伝子であるFIP1を発見した. このタンパク質は,ポリア) ポリメラーゼ (PAP1) と相互作用し,mRNAの安定性および酵母における適切な3'-end処理を維持するために不可欠です.
科学分野:
- 分子生物学は分子生物学である.
- イースト遺伝学 イースト遺伝学
- RNA 処理 RNA 処理
背景:
- ポリアデニレーションは,真核のmRNA処理における重要なステップです.
- ポリア) ポリメラーゼ (PAP1) は,ポリア) 尾を追加する酵素です.
- PAP1の活性と基質の特異性を調節する要因は完全に理解されていません.
研究 の 目的:
- イーストmRNAポリアデニレーションに関与する新しい要因を特定し,特徴づけること.
- ポリアデニレーション機械の相互作用ネットワークを解明する.
- ポリアデニレーションプロセスにおけるFIP1の役割を理解するために.
主な方法:
- タンパク質の相互作用を特定するための2つのハイブリッドアッセイ.
- 複合体の形成に関するインビトロアッセイは,再結合タンパク質を用いて行われます.
- 熱感性FIP1アレルの分析.
- イースト抽出物を用いたインビトロ3'-エンド加工アッセイ.
- コイムノプレシピテーション実験.
主要な成果:
- エッセンシャル遺伝子であるFIP1は,酵母PAP1と相互作用するタンパク質をコードする.
- FIP1は,分裂因子I (CF I) のサブユニットであるRNA14と直接相互作用する.
- FIP1の突然変異により,ポリー (A) テイルが短くなり,トランスクリプトレベルが低下する.
- FIP1ミュータント抽出物は,ポリアデニレーションの欠陥を示しますが,割れにはなりません.
- ポリアデニル化活動は,ポリアデニル化因子I (PF I) によって回復されます.
結論:
- FIP1は酵母ポリアデニレーション機構の重要な構成要素です.
- FIP1は,PAP1とCFIをRNA14経由で結びつけるブリッジとして機能します.
- この相互作用は,プレ-mRNAに対してPAP1に基板特異性を与える可能性が高い.
- FIP1は,mRNAの処理と安定性を調節する上で重要な役割を果たします.
関連する概念動画
02:01pre-mRNA Processing
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
02:35Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
02:41Chromatin Structure Regulates pre-mRNA Processing
In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
The chromatin structure, especially...
02:57piRNA - Piwi-interacting RNAs
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02:35Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
01:35Pre-mRNA Processing: Modification of pre-mRNA Ends
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...