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Restarting Stalled Replication Forks

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,...

Restarting Stalled Replication Forks

Restarting Stalled Replication Forks

The DNA Replication Fork

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...

The DNA Replication Fork

The DNA Replication Fork

DNA Replication

DNA Replication

DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied. After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication...

Chromosome Replication

Chromosome Replication

Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins. This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin...

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関連する実験動画

Updated: Feb 13, 2026

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

複製フォークの進行,保護,再起動を監視するための定量DNAファイバーアッセイ.

Debanjali Bhattacharya¹, Ganesh Nagaraju¹

¹Department of Biochemistry, Indian Institute of Science, Bangalore, India.

|February 12, 2026

まとめ

この要約は機械生成です。

この研究では,単一分子解像度でDNA複製ダイナミクスを視覚化するためのDNAファイバーアッセイを導入しています. この方法は,様々な細胞ストレス下での複製パラメータに関する定量的な洞察を提供し,以前の技術の限界を克服します.

キーワード:

DNA繊維測定法によるDNA繊維測定法フォークアシンメトリー・アシメトリーフォーク・プロテクション (フォーク・プロテクション)フォーク再起動フォークスピードはフォークスピードです.レプリケーションレプリケーションレプリケーション複製のストレスが重なる.

さらに関連する動画

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Published on: February 3, 2022

関連する実験動画

Last Updated: Feb 13, 2026

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique

Published on: October 27, 2011

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Detection of Post-Replicative Gaps Accumulation and Repair in Human Cells Using the DNA Fiber Assay

Published on: February 3, 2022

科学分野:

分子生物学は分子生物学である.
遺伝学遺伝学とは
細胞生物学細胞生物学

背景:

DNA複製は細胞分裂に不可欠ですが,様々なストレスに直面します.
複製のダイナミクスを研究する以前の方法は,空間的解像度が限られ,平均的な見積もりを提供していました.
生理学的およびストレス条件下での複製を理解することは,ゲノムの安定性にとって極めて重要です.

研究の目的:

DNA複製ダイナミクスの高解像度可視化のための新しいDNAファイバーアッセイを記述する.
単一分子レベルで複製パラメータの定量分析を可能にする.
正常およびストレス条件下での複製を研究するための再現可能な方法を提供すること.

主な方法:

複製DNAにチミジンアナログ (CldUとIdU) を組み込む.
グラススライドにラベルを貼ったDNAを伸縮して固定し,その後デナチュレーションを行う.
CldU/IdUに対する抗体と光顕微鏡を用いて,ラベル付けされた新生DNAの可視化.

主要な成果:

DNAファイバーアッセイは,単一分子解像度で個々の複製DNA分子を視覚化することを可能にします.
この技術は定量的で,高通量であり,異なる細胞系で再現可能である.
DNA合成速度,フォーク保護,再起動,複製ストレス下での非対称性についての洞察を提供します.

結論:

DNAファイバーアッセイは,DNA複製のダイナミクスを研究するための強力で汎用的なツールです.
古い方法よりも大きな利点があり,複製メカニズムに関する詳細な洞察を提供します.
このテクニックは,遺伝子毒性物質と細胞ストレスがゲノム複製に及ぼす影響を調査するのに価値があります.