Genome-wide chemical mapping of O-GlcNAcylated proteins in Drosophila melanogaster
Genome-wide chemical mapping of O-GlcNAcylated proteins in Drosophila melanogaster
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DOI：
10.1038/nchembio.2247
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发表时间：
2017-02-01
影响因子：
14.8
通讯作者：
Vocadlo, David J.
中科院分区：
文献类型：
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作者：
Liu, Ta-Wei;Myschyshyn, Mike;Vocadlo, David J.
N-乙酰葡糖胺β-O-连接核质蛋白（O-GlcNAc）参与生物体（从人类到果蝇）基因表达的调节。在果蝇中，O-GlcNAc转移酶（OGT）是Polycomb组蛋白（PcG）中的一种，其通过Polycomb组反应元件（PRE）作用于同源异型（HOX）和其他PcG靶基因。使用果蝇，我们确定了新的O-GlcNAc酰化PcG蛋白，并开发了一种无抗体的代谢喂养方法，以化学选择性地映射富含O-GlcNAc的基因组位点，使用下一代测序。我们发现O-GlcNAc在细胞和体内均分布于特定的基因组位点。这些基因座中的许多与PRE重叠，但O-GlcNAc也存在于其他缺乏PRE的基因座中。OGT的缺失不仅导致含有PRE的基因座的基因表达改变，而且导致缺乏PRE的基因座的基因表达改变，包括几个异染色质基因。这些数据表明，O-GlcNAc通过多种机制来调节果蝇中的基因表达。
N-Acetylglucosamine beta-O-linked to nucleocytoplasmic proteins (O-GlcNAc) is implicated in the regulation of gene expression in organisms, from humans to Drosophila melanogaster. Within Drosophila, O-GlcNAc transferase (OGT) is one of the Polycomb group proteins (PcGs) that act through Polycomb group response elements (PREs) to silence homeotic (HOX) and other PcG target genes. Using Drosophila, we identify new O-GlcNAcylated PcG proteins and develop an antibody-free metabolic feeding approach to chemoselectively map genomic loci enriched in O-GlcNAc using next-generation sequencing. We find that O-GlcNAc is distributed to specific genomic loci both in cells and in vivo. Many of these loci overlap with PREs, but O-GlcNAc is also present at other loci lacking PREs. Loss of OGT leads to altered gene expression not only at loci containing PREs but also at loci lacking PREs, including several heterochromatic genes. These data suggest that O-GlcNAc acts through multiple mechanisms to regulate gene expression in Drosophila.