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Deciphering the Dynamics of UHRF1‐dependent DNA Methylation
Author(s) -
Tiedemann Rochelle L.,
Jones Peter A.,
Rothbart Scott B.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.595.17
Subject(s) - dna methylation , epigenome , epigenomics , biology , epigenetics , chromatin , histone , genetics , computational biology , reprogramming , microbiology and biotechnology , dna , gene , gene expression
The epigenetic inheritance of DNA methylation patterning is a coordinated process involving UHRF1‐directed recruitment of DNMT1 to S‐phase chromatin, where the parental methylation patterns are copied on nascent DNA. As UHRF1 is now recognized as a key regulator of DNA methylation maintenance, it is considered as a promising therapeutic target for modulating the DNA methylation program in cancer and other diseases. UHRF1 itself is targeted to chromatin in a multivalent manner by the interconnected activities of three reader domains, which directs its ubiquitin ligase activity towards several lysine residues on histone H3 (a proposed docking site for DNMT1). While the mechanistic links between UHRF1 and DNA methylation maintenance are beginning to be unraveled, little is known about the dynamics of this process at epigenome‐scale resolution. To study the dynamics of UHRF1‐dependent DNA methylation, we engineered a human colorectal carcinoma cell line that expresses a doxycycline‐inducible shRNA targeting UHRF1. Importantly, this system is reversible, allowing us to model the effects of acute depletion of UHRF1 in these cells. Using this system, we are following UHRF1‐dependent DNA methylation dynamics using Illumina's Infinium Methylation EPIC Bead Array and layering this information with transcriptome, nucleosome occupancy, and histone modification dynamics. We are also querying the contributions of specific UHRF1 regulatory domains to the maintenance of epigenome integrity. Collectively, the results of this study are providing high‐resolution temporal snapshots of epigenetic programming (and reprogramming) facilitated by UHRF1 and are revealing how the epigenome responds to targeted inhibition of UHRF1 function. Support or Funding Information NIH NCI R00 CA181343 to SBR; NIH NCI R01 CA082422, R01 CA170550 to PAJ