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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,...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

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Translesion DNA Polymerases

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TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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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...

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Updated: Jul 26, 2025

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

ブレイク誘発複製は切除依存のテンプレートスイッチングをオーケストラする.

Tianpeng Zhang¹, Yashpal Rawal², Haoyang Jiang¹

¹Department of Cancer Biology, Penn Center for Genome Integrity, Basser Center for BRCA, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.

|June 14, 2023

まとめ

この要約は機械生成です。

断裂誘発テロメア合成 (BITS) は,DNA修復のために最小のレプリソームを使用します. SNM1A核酵素は,この過程でリセクションと病変バイパスを促進するために,ユビキチン化されたPCNAを誘導する.

さらに関連する動画

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

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Published on: April 28, 2021

関連する実験動画

Last Updated: Jul 26, 2025

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

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

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Assessment of Global DNA Double-Strand End Resection using BrdU-DNA Labeling coupled with Cell Cycle Discrimination Imaging

Published on: April 28, 2021

科学分野:

分子生物学
DNA 修復
テロメア生物学

背景:

断裂誘発テロメア合成 (BITS) は,テロメアの代替延長に不可欠なRAD51独立経路である.
BITSは,広範囲なDNA修復合成のために,増殖細胞核抗原 (PCNA) とDNAポリメラーゼ-δを含む最小の複素体を含みます.
複製ストレスと二次DNA構造に対するBITSの反応は十分に理解されていません.

研究の目的:

複製ストレス下でのBITS中のDNA損傷反応を調査する.
長い経路の同型再結合修復中にプロセス性維持に関与する主要なタンパク質を特定する.
BITSが複雑なDNA構造を許容するメカニズムを解明する.

主な方法:

シンクロン・インダクション・ダブル・ストランド・ブレイク
テロメア DNA 損傷反応プロテオームを分析するために,分離されたクロマチンのセグメント (PICh) のプロテオミクス.
RAD18依存のPCNAユビキチン化とSNM1Aヌクレアース活性に関する分析.

主要な成果:

BITSは,RAD18依存のPCNAユビキチネーションによって特徴づけられる複製ストレス反応を誘発する.
SNM1A核酵素は,ユビキチン化PCNA依存性DNA損傷耐性の重要なエフェクタとして特定されています.
SNM1Aは,ブレイク誘発のレプリソームでユビキチン化されたPCNAを認識し,その核酵素活性を誘導して切除を促進します.

結論:

断裂誘発複製は切除に依存する病変バイパスをオーケストラします.
哺乳類の細胞におけるユビキチン化PCNA誘導再結合には,SNM1A核酵素の活性が不可欠である.
この研究は,BITSでDNA損傷耐性の新しいメカニズムを明らかにしています.