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Published on: June 12, 2018
A method for purification, identification and validation of DNMT1 mRNA binding proteins
Alexander Unterberger1, Jérôme Torrisani, Moshe Szyf
1McGill University, 3655, Sir William-Osler Promena, Montreal, QC, Canada.
Biological Procedures Online
|December 3, 2008
Summary
AUF 1 protein binds to DNA methyltransferase 1 mRNA, triggering its destabilization during the G(o)/G(1) phase. This identifies AUF 1 as a key regulator of DNA methyltransferase 1 (DNMT1) expression.
Area of Science:
- Molecular Biology
- Epigenetics
- Gene Regulation
Background:
- DNA methyltransferase 1 (DNMT1) maintains DNA methylation patterns crucial for cell division.
- DNMT1 expression is tightly regulated throughout the cell cycle.
- Previous research indicated a ~40 kDa protein binding to DNMT1 3'-UTR destabilizes its mRNA during G(o)/G(1) phase.
Purpose of the Study:
- To identify the protein binding to the DNMT1 3'-UTR.
- To validate the role of the identified protein in DNMT1 mRNA destabilization.
Main Methods:
- RNA affinity capture using DNMT1 3'-UTR.
- Matrix-assisted laser desorption-time of flight tandem mass spectrometry (MALDI-TOF-MS-MS) for protein identification.
- Validation experiments to confirm protein function.
Main Results:
- AUF 1 (AU-rich element binding factor 1) was identified as the protein binding to DNMT1 mRNA.
- AUF 1 binding was confirmed to destabilize DNMT1 mRNA transcripts.
- This mechanism regulates DNMT1 expression during the G(o)/G(1) phase.
Conclusions:
- AUF 1 is the protein responsible for the destabilization of DNMT1 mRNA.
- AUF 1 plays a critical role in the cell cycle-specific regulation of DNMT1 expression.
- This finding provides insight into the post-transcriptional control of epigenetic maintenance.
