Over the past several years, we have attempted to derive a greater understanding of the role of 5-hydroxymethylcytosine (5hmC) modified DNA in mammalian DNA. Initially, we showed that 5hmC could inhibit the activity of RNA polymerase II by preventing this enzyme from binding to the promoter. The presence of 5hmC within the gene body has little effect on the ability of RNA polymerase II from elongating or terminating transcripts. This finding is important, as most 5hmC has been shown by others to be located within the gene body and to a limited degree at promoters and enhancer regions.
More recently, we developed a method for the efficient and selective identification of DNA containing the 5hmC modification. This method takes advantage of two proteins: the T4 β-glucosyltransferase, which converts 5hmC to β-glucosyl-5hmC; and JBP1, which specifically recognizes and binds to β-glucosyl-5hmC.
We have identified several proteins that specifically interact with 5hmC-modified DNA. We have found proteins that appear to modulate the transcription of specific 5hmC-modified promoters. In this way it appears that 5hmC regulates transcription by recruiting proteins that either inhibit or enhance transcription. We are interested in DNA transactions that involve 5hmC. With this in mind we have created and are in the process of creating more transgenic mouse models that will allow us to evaluate these unique DNA transactions.
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