5-Hydroxymethylcytosine Is Not Present in Appreciable Quantities in Arabidopsis DNA
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Gehring_5 hydroxy.pdf
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Author(s)
Gehring, Mary
Erdmann, Robert Michael
Souza, Amanda L.
Clish, Clary
Date Issued
November 2014
Journal
G3: Genes-Genomes-Genetics
Publisher
Genetics Society of America, The
Citation
Erdmann, Robert M., Amanda L. Souza, Clary B. Clish, and Mary Gehring. “5-Hydroxymethylcytosine Is Not Present in Appreciable Quantities in Arabidopsis DNA.” G3: Genes-Genomes-Genetics (November 6, 2014).
Version
Final published version
Abstract
5-Hydroxymethylcytosine (5-hmC) is an intermediate in active demethylation in metazoans, as well as a potentially stable epigenetic mark. Previous reports investigating 5-hydroxymethylcytosine in plants have reached conflicting conclusions. We systematically investigated whether 5-hmC is present in plant DNA using a range of methods. Using the model organism Arabidopsis thaliana, in addition to other plant species, we assayed the amount or distribution of 5-hydroxymethylcytosine by thin-layer chromatography, immunoprecipitation-chip, ELISA, enzymatic radiolabeling, and mass spectrometry. The failure to observe 5-hydroxymethylcytosine by thin-layer chromatography established an upper bound for the possible fraction of the nucleotide in plant DNA. Antibody-based methods suggested that there were low levels of 5-hmC in plant DNA, but these experiments were potentially confounded by cross-reactivity with the abundant base 5-methylcytosine. Enzymatic radiolabeling and mass spectrometry, the most sensitive methods for detection that we used, failed to detect 5-hydroxymethylcytosine in A. thaliana genomic DNA isolated from a number of different tissue types and genetic backgrounds. Taken together, our results led us to conclude that 5-hmC is not present in biologically relevant quantities within plant genomic DNA.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Whitehead Institute for Biomedical Research
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Creative Commons Attribution Unported License
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DOI of Published Version
https://doi.org/10.1534/g3.114.014670
