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Related Concept Videos

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...
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...
The DNA Replication Fork01:02

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 forks, one in...
The DNA Replication Fork01:02

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 forks, one in...
The Spindle Assembly Checkpoint02:19

The Spindle Assembly Checkpoint

The spindle assembly checkpoint is a molecular surveillance mechanism ensuring the fidelity of chromosome segregation during anaphase. The checkpoint monitors the completion of all the prerequisite steps before chromosome segregation to determine whether the segregation process should proceed or be delayed.
Many proteins function together to control the spindle assembly checkpoint. Mutations affecting these proteins may allow cells to proceed into anaphase prematurely, resulting in the...
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.

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Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
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Published on: October 27, 2011

Claspin promotes normal replication fork rates in human cells.

Eva Petermann1, Thomas Helleday, Keith W Caldecott

  • 1Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton BN1 9RQ, United Kingdom. eva.petermann@rob.ox.ac.uk

Molecular Biology of the Cell
|March 21, 2008
PubMed
Summary

Claspin is essential for rapid DNA replication fork progression in human cells, similar to Chk1. Depleting Claspin slows replication forks and may have roles beyond Chk1 phosphorylation.

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

Published on: February 3, 2022

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Claspin (S phase-specific adaptor protein) and Chk1 are key components of the ATR pathway, crucial for responding to replication stress.
  • Evidence indicates these ATR pathway components also play vital roles during normal S phase DNA replication.

Purpose of the Study:

  • To investigate the role of human Claspin in regulating replication fork progression during normal S phase.
  • To determine if Claspin is required for maintaining high replication fork rates in human cells.

Main Methods:

  • DNA fiber labeling techniques were employed in HeLa, HCT116, and primary human 1BR3 fibroblast cells.
  • Depletion of Claspin and/or Chk1 was performed to assess effects on replication fork speed.

Main Results:

  • Depletion of Claspin significantly slowed replication fork progression in all tested human cell types, comparable to Chk1 depletion.
  • Claspin-depleted cells still showed Chk1 phosphorylation, suggesting Claspin's role may exceed facilitating this specific phosphorylation.
  • Combined depletion of Chk1 and Claspin exacerbated replication fork slowing more than individual depletions.

Conclusions:

  • Human Claspin is universally required for maintaining high replication fork progression rates during normal S phase.
  • Claspin's function in normal DNA replication may involve mechanisms independent of facilitating Chk1 phosphorylation at Ser317/Ser345.
  • Synergistic roles of Claspin and Chk1 are critical for preventing severe replication fork slowing.