Related Experiment Video
Updated: May 28, 2025
08:53
Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
Published on: May 2, 2025
187
Identification of MCM2-Interacting Proteins Associated with Replication Initiation Using APEX2-Based Proximity
Sitong Yao1,2, Zhen Yue2, Shaotang Ye1
1College of Veterinary Medicine, South China Agricultural University, 483 Wushan Road, Guangzhou 510642, China.
International Journal of Molecular Sciences
|February 13, 2025
Summary
Researchers identified new proteins interacting with the MCM2 replication initiation complex using APEX2 labeling and mass spectrometry. This advances understanding of DNA replication and genomic stability.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- DNA replication is vital for cell growth and accurate genetic transmission.
- Replication initiation involves the pre-replication complex (pre-RC), but transient interactions are hard to detect.
- MCM2 is a key component of the eukaryotic replication initiation complex.
Purpose of the Study:
- To identify proteins proximal to the MCM2 replication initiation complex.
- To investigate transient protein interactions in DNA replication initiation.
- To understand the regulatory network of the DNA replication initiation complex.
Main Methods:
- Fused MCM2 with enhanced APEX2 for in situ biotinylation.
- Employed mass spectrometry to identify proximal proteins in mouse ESCs and NIH/3T3 cells.
- Confirmed interactions using bimolecular fluorescence complementation (BiFC).
Main Results:
- Identified candidate proteins proximal to the MCM2 complex.
- Confirmed interactions between MCM2 and CD2BP2, VRK1, and GTSE1.
- Established a method for detecting transient protein interactions in replication initiation.
Conclusions:
- The study identified novel candidate proteins involved in DNA replication initiation.
- This research provides a foundation for studying the conserved DNA replication initiation complex and its regulatory network.
- The APEX system is effective for detecting transient protein interactions in complex biological processes.
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