Immunoprecipitation and mass spectrometry define TET1 interactome during oligodendrocyte differentiation
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Cell & Bioscience Open Access
RESEARCH
Immunoprecipitation and mass spectrometry define TET1 interactome during oligodendrocyte differentiation Ming Zhang1, Kaixiang Zhang1, Jian Wang1, Yuming Liu1, Guangxin Liu1, Weilin Jin2, Shengxi Wu1* and Xianghui Zhao1*
Abstract Ten-eleven translocation (TET) proteins, encoding dioxygenase for DNA hydroxymethylation, are important players in nervous system development and disease. In addition to their proverbial enzymatic role, TET proteins also possess non-enzymatic activity and function in multiple protein–protein interaction networks, which remains largely unknown during oligodendrocyte differentiation. To identify partners of TET1 in the myelinating cells, we performed proteome-wide analysis using co-immunoprecipitation coupled to mass spectrometry (IP-MS) in purified oligodendrocyte precursor cells (OPCs) and mature oligodendrocytes (mOLs), respectively. Following a stringent selection of MS data based on identification reliability and protein enrichment, we identified a core set of 1211 partners that specifically interact with TET1 within OPCs and OLs. Analysis of the biological process and pathways associated with TET1interacting proteins indicates a significant enrichment of proteins involved in regulation of cellular protein localization, cofactor metabolic process and regulation of catabolic process, et al. We further validated TET1 interactions with selected partners. Overall, this comprehensive analysis of the endogenous TET1 interactome during oligodendrocyte differentiation suggest its novel mechanism in regulating oligodendrocyte homeostasis and provide comprehensive insight into the molecular pathways associated with TET1. Keywords: Oligodendrocyte, TET1, DNA dioxygenase, Mass spectrometry, Co-immunoprecipitation Introduction Emerging as new epigenetic factors for neural development, TET family members have been associated with the process of oligodendrocyte (OL) differentiation [1]. Axon myelination by OLs enables saltatory conduction of action potentials and provides long-term trophic support for axons, maintaining integrity throughout the central nervous system (CNS) [2]. TET proteins were initially identified as dioxygenase for DNA hydroxymethylation and oxidize 5-methylcytosin (5mC) to *Correspondence: [email protected]; [email protected] 1 Department of Neurobiology, School of Basic Medicine, Fourth Military Medical University, Xi’an 710032, Shaanxi, China Full list of author information is available at the end of the article
5-hydroxymethylcytosine (5hmC) to initiate the DNA demethylation process. Three members of the mammalian TET gene family have been defined, TET1, 2 and 3, and all TET proteins contain a similar C-terminal catalytic domain, which confers α-ketoglutarate (α-KG)-and iron (II)-dependent dioxygenase activity [3]. Extensive studies have focused on the catalytic enzymatic roles of TET family proteins in regulating various process from development to disease in a cell-type and context-dependent manner [4]; while some investiga
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