Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
Methods Enzymol. 2012;512:93-105. doi: 10.1016/B978-0-12-391940-3.00005-6.

Enzymatic analysis of Tet proteins: key enzymes in the metabolism of DNA methylation.

Author information

  • 1Howard Hughes Medical Institute; Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

Abstract

One of the most exciting recent advances in the epigenetic field is the discovery that 5-methylcytosine (5mC) in DNA can be iteratively oxidized by a family of proteins known as Tet proteins to generate 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). These 5mC derivatives can be further processed by thymine-DNA glycosylase (TDG) followed by base excision repair or by replication-dependent dilution leading to DNA demethylation. Given the similarity between 5mC and its oxidation derivatives, many of the conventional techniques used for 5mC analysis cannot distinguish between 5mC and 5hmC/5fC/5caC. Here, we describe 2D-TLC and mass spectrometry methods that we have successfully used in differentiating 5mC from its oxidative derivatives as well as in characterizing the enzymatic activity of Tet proteins both in vitro and in vivo.

Copyright © 2012 Elsevier Inc. All rights reserved.

PMID:
22910204
[PubMed - indexed for MEDLINE]
PMCID:
PMC3555433
Free PMC Article

Images from this publication.See all images (3)Free text

Figure 5.1
Figure 5.2
Figure 5.3
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk