5-hydroxymethylcytosine (5hmC) has been suggested to be involved in various nucleic acid transactions and cellular processes, including transcriptional regulation, demethylation of 5-methylcytosine and stem cell pluripotency. An activity has been identified that preferentially catalyzes the cleavage of double-stranded 5hmC-modified DNA. Using biochemical methods this activity has been purified from mouse liver extracts and demonstrate that the enzyme responsible for the cleavage of 5hmC-modified DNA is Endonuclease G (EndoG). It has been show that recombinant EndoG preferentially recognizes and cleaves a core sequence when one specific cytosine within that core sequence is hydroxymethylated. Additionally, in vivo evidence has been provided that EndoG catalyzes the formation of double-stranded DNA breaks and that this cleavage is dependent upon the core sequence, EndoG and 5hmC. Finally, it has been demonstrated that the 5hmC modification can promote conservative recombination in an EndoG-dependent manner.
Mouse EndoGSKU: P-288
About the scientist
My research group has two principal research foci: (i) studying the cellular processes involving in the metabolism of the modified cytosine base – 5-hydroxymethylcytosine and (ii) the development of a continuous directed evolution platform that can be used in eukaryotic cells.