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Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Stringent Response in E. coli01:23

Stringent Response in E. coli

Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

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Updated: Jun 25, 2026

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
10:15

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

Published on: July 6, 2012

単一ラムダ外核酵素分子の配列依存的な停止.

Thomas T Perkins1, Ravindra V Dalal, Paul G Mitsis

  • 1Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA. tperkins@jila.colorado.edu

Science (New York, N.Y.)
|August 30, 2003
PubMed
まとめ
この要約は機械生成です。

ラムダ外核酵素は,DNA鎖を一定速度で消化しますが,特定のDNA配列で一時停止します. これらの一時停止は,特にGGCGAモチーフでは,DNA再結合効率に影響を与える可能性があります.

さらに関連する動画

Electricity-Free, Sequential Nucleic Acid and Protein Isolation
09:52

Electricity-Free, Sequential Nucleic Acid and Protein Isolation

Published on: May 15, 2012

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution
10:27

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution

Published on: July 8, 2019

関連する実験動画

Last Updated: Jun 25, 2026

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA
10:15

Isolation of Ribosome Bound Nascent Polypeptides in vitro to Identify Translational Pause Sites Along mRNA

Published on: July 6, 2012

Electricity-Free, Sequential Nucleic Acid and Protein Isolation
09:52

Electricity-Free, Sequential Nucleic Acid and Protein Isolation

Published on: May 15, 2012

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution
10:27

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution

Published on: July 8, 2019

科学分野:

  • 分子生物学は分子生物学である.
  • バイオケミストリー バイオケミストリー
  • 遺伝学 遺伝学とは

背景:

  • ラムダエキゾヌクレアゼは,DNA代謝における重要な酵素である.
  • 単一分子レベルでその酵素活性を理解することは,DNA処理の洞察を提供します.
  • 以前の研究では,その一般的過程性および方向性を特徴づけた.

研究 の 目的:

  • ラムダ外核酵素消化の単分子運動を調査する.
  • DNA分解中のシーケンス固有の停止イベントを特定するために.
  • DNA配列と潜在的な生物学的影響と一時停止を相関させる.

主な方法:

  • ラムダ外核酵素の活性に関する単分子観察.
  • DNAの消化速度と一時停止期間を測定する.
  • 強い停止配列を特定するために,ゲル電泳.
  • 特定のモチーフと一時停止を相関させるためのシーケンス分析.

主要な成果:

  • ラムダ外核酵素は,ほぼ恒常速度 (4 nm/s) で過程的な分解を示します.
  • 消化過程は,特定の場所において,期間が変動するパウズによって断続される.
  • パウスは鎖特異的でシーケンスに依存し,GGCGAモチーフで強いパウスが識別されます.
  • GGCGAモチーフは,左のラムダ結束端にある.

結論:

  • 単一分子分析は,配列依存の一時停止を含む,ラムダ外核酵素の微妙な動力学を明らかにします.
  • 特定されたGGCGAモチーフは,重要なパウズサイトとして機能します.
  • このモチーフにおけるエキゾヌクレアース阻害は,ラムダ結束端での再結合効率の低下に貢献する可能性があります.