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Quantitative proteomics revealed new functions of ALKBH4
Authors:Kailin Yu  Tianyu F Qi  Weili Miao  Xiaochuan Liu  Yinsheng Wang
Institution:1. Department of Chemistry, University of California, Riverside, California, USA;2. Environmental Toxicology Graduate Program, University of California, Riverside, California, USA
Abstract:ALKBH4 is a versatile demethylase capable of catalyzing the demethylation of monomethylated lysine-84 on actin and N6-methyladenine in DNA. In this study, we conducted a quantitative proteomic experiment to reveal the altered expression of proteins in HEK293T cells upon genetic ablation of ALKBH4. Our results showed markedly diminished levels of GSTP1 and HSPB1 proteins in ALKBH4-depleted cells, which emanate from an augmented expression level of DNA (cytosine-5)-methyltransferase 1 (DNMT1) and the ensuing elevated cytosine methylation in the promoter regions of GSTP1 and HSPB1 genes. Together, our results revealed a role of ALKBH4 in modulating DNA cytosine methylation through regulating the expression level of DNMT1 protein.
Keywords:bisulfite sequencing  cytosine methylation  mass spectrometry  parallel-reaction monitoring  post-translational modification  quantitative proteomics  SILAC
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