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Updated: Jun 7, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

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Specific origin selection and excess functional MCM2-7 loading in ORC-deficient cells.

Yoshiyuki Shibata, Mihaela Peycheva, Etsuko Shibata

Biorxiv : the Preprint Server for Biology

|November 18, 2024

Summary

This summary is machine-generated.

The six-subunit Origin Recognition Complex (ORC) is not essential for DNA replication origin specification or MCM2-7 loading in human cancer cells. Specific origins are still used, and excess MCM2-7 is loaded, even without ORC.

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Determination of the Optimal Chromosomal Locations for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Determination of the Optimal Chromosomal Locations for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Published on: September 11, 2017

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

The Origin Recognition Complex (ORC) is a six-subunit complex crucial for initiating DNA replication.
ORC is believed to play a key role in specifying where DNA replication begins on chromosomes.
Excess loading of MCM2-7 proteins is essential for licensing origins of replication.

Purpose of the Study:

To investigate the necessity of the six-subunit ORC for origin specification.
To determine if ORC is required for the loading of excess MCM2-7 proteins.
To understand the role of ORC in DNA replication initiation in human cancer cell lines.

Main Methods:

Engineered cancer cell lines with deletions in specific ORC subunits (ORC1, ORC2, ORC5).
Mapped DNA replication origins in these engineered cell lines.
Analyzed MCM2-7 loading rates and licensing of origins in G1 and S phases.

Main Results:

Specific DNA replication origins were still utilized in ORC-deficient cells, often at conserved genomic locations.
GC/TA skewness and simple repeat sequences were found to facilitate, but not dictate, origin selection without ORC.
Excess MCM2-7 was loaded at comparable rates in G1 phase and re-loaded in S phase, despite the absence of the six-subunit ORC.

Conclusions:

Origin specification during DNA replication does not require the six-subunit ORC in human cancer cell lines.
Excess MCM2-7 loading and licensing of dormant origins are independent of the six-subunit ORC.
These findings challenge the established role of ORC in origin selection and MCM loading.