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Related Concept Videos

The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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...
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
DNA Damage Can Stall the Cell Cycle02:36

DNA Damage Can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...

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Related Experiment Video

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Study of the DNA Damage Checkpoint using Xenopus Egg Extracts
10:55

Study of the DNA Damage Checkpoint using Xenopus Egg Extracts

Published on: November 5, 2012

Xenopus ATR is a replication-dependent chromatin-binding protein required for the DNA replication checkpoint.

M Hekmat-Nejad1, Z You, M C Yee

  • 1Department of Molecular Pharmacology, Stanford University, Palo Alto, California 94305-5174, USA.

Current Biology : CB
|January 4, 2001
PubMed
Summary

The ATR kinase is essential for the DNA replication checkpoint, ensuring cells do not enter mitosis before DNA synthesis is complete. Its depletion causes premature mitosis, highlighting its critical role in cell cycle regulation.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • The DNA replication checkpoint prevents premature mitosis until DNA synthesis is complete.
  • ATR kinase is a potential upstream regulator of this checkpoint, but a suitable system for study was lacking.

Purpose of the Study:

  • To investigate the function of ATR kinase in the DNA replication checkpoint using Xenopus egg extracts.
  • To characterize ATR's role in relation to DNA replication and Chk1 phosphorylation.

Main Methods:

  • Cloning of Xenopus laevis ATR (XATR).
  • Chromatin-binding assays in Xenopus egg extracts.
  • Analysis of Cdc2-cyclin B activity and Chk1 phosphorylation upon ATR depletion.

Main Results:

  • ATR associates with chromatin during replication and accumulates when replication is inhibited.
  • ATR's chromatin association depends on RNA primer synthesis by DNA polymerase alpha.
  • Depletion of ATR caused premature mitosis and reduced Chk1 phosphorylation, even with replication inhibitors.

Conclusions:

  • ATR is a replication-dependent chromatin-binding protein crucial for the DNA replication checkpoint.
  • ATR's function is linked to RNA synthesis by DNA polymerase alpha.
  • ATR is required to prevent premature entry into mitosis.