有哪些信誉好的足球投注网站a silac-based screen for methyl-cpg binding proteins identifies rbp-j as a dna methylation and sequence-specific binding proteinsilac-based屏幕methyl-cpg结合蛋白识别rbp-j dna甲基化和sequence-specific结合蛋白.pdfVIP
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a silac-based screen for methyl-cpg binding proteins identifies rbp-j as a dna methylation and sequence-specific binding proteinsilac-based屏幕methyl-cpg结合蛋白识别rbp-j dna甲基化和sequence-specific结合蛋白
A SILAC-Based Screen for Methyl-CpG Binding Proteins Identifies RBP-J as a DNA Methylation and Sequence- Specific Binding Protein 1. 2. 1 2 Stefanie J. J. Bartels , Cornelia G. Spruijt , Arie B. Brinkman , Pascal W. T. C. Jansen , Michiel 2 1 Vermeulen , Hendrik G. Stunnenberg * 1 Department of Molecular Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, The Netherlands, 2 Department of Molecular Cancer Research, University Medical Center Utrecht, Utrecht, The Netherlands Abstract Background: DNA methylation is an epigenetic modification that plays a crucial role in a variety of biological processes. Methylated DNA is specifically bound by Methyl-CpG Binding Proteins (MBPs). Three different types of MBPs have been identified so far: the Methyl-CpG Binding Domain (MBD) family proteins, three BTB/POZ-Zn-finger proteins, and UHRF1. Most of the known MBPs have been identified via homology with the MBD and Zn-finger domains as present in MeCP2 and Kaiso, respectively. It is conceivable that other proteins are capable of recognizing methylated DNA. Methodology/Principal Findings: For the purpose of identifying novel ‘readers’ we set up a methyl-CpG pull-down assay combined with stable-isotope labeling by amino acids in cell culture (SILAC). In a methyl-CpG pull-down with U937 nuclear extracts, we recovered several known MBPs and almost all subunits of the MBD2/NuRD complex as methylation specific binders, providing proof-of-principle. Interestingly, RBP-J, the transcription factor downstream of Notch receptors, also bound the DNA in a methylation dependent manner. Follow-up pull-downs and electrophoreti
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