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Updated: Apr 19, 2026

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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DNA-protein interaction dynamics at the Lamin B2 replication origin.

Luca Puzzi1, Laura Marchetti, Fiorenzo A Peverali

  • 1a Molecular Biology Laboratory; Scuola Normale Superiore ; Pisa , Italy.

Cell Cycle (Georgetown, Tex.)
|December 9, 2014
PubMed
Summary

Researchers identified AP-1 transcription factors (c-Fos and c-Jun) binding to the Lamin B2 DNA replication origin in human cells. DNA topology, influenced by topoisomerase II, is crucial for origin activation.

Keywords:
AP-1DNA replication originFosJunorigin recognition complextranscription factor

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Understanding DNA replication origin activation in mammals is incomplete.
  • The molecular mechanisms governing the initiation of DNA replication remain a key area of research.

Purpose of the Study:

  • To investigate protein interactions at the human Lamin B2 replication origin.
  • To elucidate the molecular events involved in mammalian DNA replication initiation.

Main Methods:

  • High-resolution chromatin immunoprecipitation (ChIP) was employed.
  • Proteins interacting with the Lamin B2 origin DNA were identified.
  • The effect of merbarone, a topoisomerase II inhibitor, on replication and protein binding was assessed.

Main Results:

  • The study identified ORC4, USF, and HOXC13 at the Lamin B2 origin.
  • Crucially, AP-1 transcription factors (c-Fos and c-Jun) were found associated with the origin DNA during late G1 phase.
  • These AP-1 factors were observed to interact with ORC4.
  • Merbarone treatment disrupted both DNA replication and AP-1 factor binding at the origin.

Conclusions:

  • AP-1 transcription factors (c-Fos and c-Jun) play a role in mammalian DNA replication origin activation.
  • The interaction between AP-1 factors and ORC4 suggests a novel regulatory mechanism.
  • DNA topology, modulated by topoisomerase II activity, is essential for origin function and activation.