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Termination of Translation01:44

Termination of Translation

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The large ribosomal subunit has several important structures essential to translation. These include the peptidyl transferase center (PTC) - which is the site where the peptide bond is formed - and a large, internal, water-filled tube through which the nascent polypeptide moves. This latter structure is called the Peptide Exit Tunnel, and it begins at the PTC and spans the body of the large ribosomal subunit. During translation, as the nascent polypeptide chain is synthesized, it passes through...
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Termination of Translation01:44

Termination of Translation

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Leaky Scanning02:28

Leaky Scanning

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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Initiation of Translation02:33

Initiation of Translation

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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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Initiation of Translation02:33

Initiation of Translation

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Improving Translational Accuracy02:07

Improving Translational Accuracy

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Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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関連する実験動画

Updated: Mar 11, 2026

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on TRO Approach
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ストップコドンが欠けているmRNAのArfA-RF2媒介による翻訳終結の構造的基礎

Paul Huter1, Claudia Müller1, Bertrand Beckert1,2

  • 1Gene Center, Department of Biochemistry and Center for integrated Protein Science Munich (CiPSM), Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.

Nature
|December 2, 2016
PubMed
まとめ

代替救済因子A (ArfA) は,断片されたmRNAに停滞した細菌のリボソームを救う. この研究は,ArfA

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Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
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関連する実験動画

Last Updated: Mar 11, 2026

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12:12

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Published on: March 12, 2017

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Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
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Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
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科学分野:

  • バクテリアのトランスレーション終結
  • リボソーム救済メカニズム
  • 分子冷凍電子顕微鏡

背景:

  • バクテリアはtmRNA,ArfA,またはArfBを使用して,ストップコドンが欠けている断片化されたmRNAに停滞したリボソームを救出します.
  • 以前の構造研究は,tmRNA-リボソームとArfB-リボソーム複合体を明らかにしましたが,ArfAのメカニズムは不明でした.

研究 の 目的:

  • ArfAが切断されたmRNAを認識し,リボソーム救出のための解放因子2 (RF2) を採用する構造的メカニズムを解明する.
  • ArfAとRF2との複合体で断片されたmRNAに停滞したEscherichia coli 70Sリボソームの構造を決定する.

主な方法:

  • クリオ電子顕微鏡 (cryo-EM) による,ArfAとRF2を用いた断片されたmRNAに停滞した70Sリボソームの再構築.
  • ArfA,RF2,およびリボソーム間の分子相互作用の構造分析.

主要な成果:

  • ArfAのC端は,小さなリボソームサブユニットのmRNAエントリーチャネルに結合し,断片されたmRNAと全長mRNAの区別を可能にします.
  • ArfAのN端はRF2の解読領域と相互作用し,RF2を停止したリボソームに誘導する.
  • ArfAは,GGQモチーフをペプチジルトランスフェラーゼセンターに配置することで,RF2の活性構造を安定させ,カノニカル終結を模倣する.

結論:

  • この構造はArfAがRF2を 停滞したリボソームにどう誘導するかを示しています
  • ArfAはアクティブなRF2コンフォームを促進し,ストップコドンなしで翻訳終了を可能にします.
  • これは,ArfA媒介のリボソーム救済経路の分子理解を提供します.