Selective oxidation of 5-methylcytosine by TET-family proteins
First Claim
1. A method for detecting 5-hydroxymethylcytosine in a nucleic acid, the method comprising contacting the nucleic acid with an enzyme and utilizing a glucose donor substrate or a glucose-derivative donor substrate that traps covalent enzyme-DNA intermediates to detect 5-hydroxymethylcytosine residues, wherein the enzyme comprises an alpha-glucosyltransferase, a beta-glucosyltransferase, or a beta-glucosyl-alpha-glucosyl-transferase.
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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.
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13 Claims
- 1. A method for detecting 5-hydroxymethylcytosine in a nucleic acid, the method comprising contacting the nucleic acid with an enzyme and utilizing a glucose donor substrate or a glucose-derivative donor substrate that traps covalent enzyme-DNA intermediates to detect 5-hydroxymethylcytosine residues, wherein the enzyme comprises an alpha-glucosyltransferase, a beta-glucosyltransferase, or a beta-glucosyl-alpha-glucosyl-transferase.
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