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Visualization of DNA Replication in the Vertebrate Model System DT40 using the DNA Fiber Technique
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TOP1 and TOP2 complementarily maintain DNA replication fork progression in vertebrates.

Koyuki Umemura1, Masato Ooka2, Miku Sojo1

  • 1Department of Chemistry, Graduate School of Science, Tokyo Metropolitan University, Tokyo, Japan.

The Journal of Biological Chemistry
|March 6, 2026
PubMed
Summary
This summary is machine-generated.

Topoisomerases TOP1 and TOP2 are crucial for DNA replication. This study reveals their complementary roles in vertebrate cells, impacting both replication progression and initiation.

Keywords:
DT40TOP1TOP2topoisomerase

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Topoisomerases TOP1 and TOP2 manage DNA topological stress during replication.
  • Their complementary functions in yeast are known, but their interplay in vertebrate cells is unclear.

Purpose of the Study:

  • To investigate the functional interplay between TOP1 and TOP2 in vertebrate DNA replication.
  • To determine if TOP1 and TOP2 exhibit complementary roles in vertebrate cells.

Main Methods:

  • Utilized conditionally TOP1-depleted chicken DT40 cells, a model for gene editing.
  • Employed TOP2 inhibitor ICRF193 and TOP2 poison etoposide.
  • Assessed replication fork progression, S phase arrest, and apoptosis.

Main Results:

  • TOP1 depletion alone or TOP2 inhibition alone did not impede DNA replication.
  • Combined TOP1 depletion and TOP2 inhibition nearly abolished DNA replication, causing S phase arrest and apoptosis.
  • TOP1 depletion combined with etoposide treatment led to G1/early S phase arrest, indicating impaired replication initiation.

Conclusions:

  • TOP1 and TOP2 possess complementary roles in vertebrate DNA replication.
  • These topoisomerases are essential for both replication progression and initiation in vertebrate cells.