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MCM3AP, a novel acetyltransferase that acetylates replication protein MCM3
Y Takei1, M Swietlik, A Tanoue
1Wellcome/CRC Institute, Cambridge, UK. yt215@cam.ac.uk
EMBO Reports
|March 22, 2001
Summary
We discovered MCM3AP, an enzyme that acetylates MCM3, a key protein in DNA replication. This acetylation process appears to regulate DNA replication, revealing a new regulatory pathway.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Minichromosome maintenance (MCM) proteins are crucial for initiating DNA replication.
- MCM3 is a core component of the MCM complex, essential for DNA replication.
- A novel MCM3-associated protein (MCM3AP) was identified.
Purpose of the Study:
- To investigate the function of MCM3AP.
- To determine if MCM3AP acetylates MCM3.
- To explore the role of MCM3 acetylation in DNA replication.
Main Methods:
- Two-hybrid screening to identify MCM3AP.
- In vitro and in vivo acetylation assays.
- Site-directed mutagenesis of putative acetyl-CoA binding motifs in MCM3AP.
- Over-expression studies of MCM3AP and its mutants.
Main Results:
- MCM3AP was identified as an acetyltransferase that specifically acetylates MCM3.
- Chromatin-bound MCM3 is acetylated in vivo, and MCM3AP is also chromatin-bound.
- Conserved acetyl-CoA binding motifs in MCM3AP are essential for its acetyltransferase activity.
- Over-expression of MCM3AP inhibits DNA replication, an effect abolished by mutating the acetylase motifs.
Conclusions:
- MCM3AP acetylates MCM3, suggesting a novel regulatory mechanism for DNA replication.
- Acetylation of MCM3 by MCM3AP appears to play a role in regulating DNA replication.
- This study reveals a new pathway involving MCM3 acetylation in DNA replication control.