000001657 001__ 1657 000001657 005__ 20250829130723.0 000001657 0247_ $$2doi$$a10.6082/uchicago.1657 000001657 037__ $$aTHESIS$$bDissertation 000001657 041__ $$aeng 000001657 245__ $$aCharacterization of oxidized methylcytosine binding protein activities in the mammalian brain and stem cells 000001657 260__ $$bUniversity of Chicago 000001657 269__ $$a2016-06 000001657 300__ $$a185 000001657 336__ $$aDissertation 000001657 502__ $$bPh.D. 000001657 520__ $$a5-Methylcytosine embedded in mammalian DNA represses local transcription by recruiting modification-specific binding partners. Its active removal is initiated by sequential oxidation of the 5-methyl group by TET enzymes to produce three oxidized species, collectively referred to as [ox]mC. Although rare, the distribution of [ox]mC modifications is tissue-, gene-, and coding strand-specific and distinct from 5-methylcytosine, suggesting unique functions. To examine this possibility, I fractionated mammalian brain extracts to discover, isolate and characterize binding partners specific for [ox]mC. This purification reveals remarkably specific factors that are selective for each of the three oxidation states and sensitive to the 5-modification state on each strand. I demonstrate that one such factor, WDR76, is a highly 5-hydroxymethylcytosine-specific binding protein. I have begun to lay the foundation for further mechanistic studies of these specific binding proteins in mouse embryonic stem cells and leukemia. My results provide an essential bridge from studies of the distribution of [ox]mC and the effects of TET knockouts, to the possible functions of [ox]mC recognition in gene regulation or chromatin signaling. 000001657 542__ $$fCC BY-NC-ND 000001657 650__ $$aBiochemistry 000001657 653__ $$aDNA methylation 000001657 653__ $$aEpigenetics 000001657 690__ $$aBiological Sciences Division 000001657 690__ $$aPritzker School of Medicine 000001657 691__ $$aBiochemistry and Molecular Biology 000001657 7001_ $$aMalecek, Kathryn Elizabeth$$uUniversity of Chicago 000001657 72012 $$aAlexander J. Ruthenburg 000001657 72014 $$aPhoebe Rice 000001657 72014 $$aSean Crosson 000001657 72014 $$aJoseph Piccirilli 000001657 8564_ $$9d5c2b80e-4a96-48dd-96d0-bb6b0f0bae0c$$eEmbargo (2018-06-20)$$s55361939$$uhttps://knowledge.uchicago.edu/record/1657/files/Malecek_uchicago_0330D_13331.pdf 000001657 909CO $$ooai:uchicago.tind.io:1657$$pDissertations$$pGLOBAL_SET 000001657 983__ $$aDissertation