Selective oxidation of 5-methylcytosine by TET-family proteins
AssigneesChildren's Medical Center Corporation, The United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsAnjana Rao, Mamta Tahiliani, Kian Peng Koh, Suneet Agarwal, Aravind Iyer
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Abstract
The present invention provides for novel methods for regulating and detecting the cytosine methylation status of DNA. The invention is based upon identification of a novel and surprising catalytic activity for the family of TET proteins, namely TET1, TET2, TET3, and CXXC4. The novel activity is related to the enzymes being capable of converting the cytosine nucleotide 5-methylcytosine into 5-hydroxymethylcytosine by hydroxylation.
Claims10
A method for distinguishing by glycosylation 5-hydroxymethylcytosine from 5-methylcytosine in a mammalian nucleic acid sequence, said method comprising:
- (a) contacting said mammalian nucleic acid sequence with an enzyme that utilizes a labeled glucose or a labeled glucose-derivative donor substrate to add a labeled glucose molecule or a labeled glucose-derivative to a 5-hydroxymethylcytosine to generate a labeled glucosylated-5-hydroxymethylcytosine,
- wherein the enzyme is a beta-glucosyltransferase and
- wherein the labeled glucose or the labeled glucose-derivative donor substrate comprises a uridine diphosphate glucose (UDPG);
- and
- (b) distinguishing by glycosylation the 5-hydroxymethylcytosine from the 5-methylcytosine by detecting the labeled glucose molecule or the labeled glucose-derivative.
Dependencies are inferred from the claim text.