O-Linked N-acetylglucosamine transferase 1 regulates global histone H4 acetylation via stabilization of the nonspecific lethal protein NSL3
O-Linked N-acetylglucosamine transferase 1 regulates global histone H4 acetylation via stabilization of the nonspecific lethal protein NSL3
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O-连接 N-乙酰氨基葡萄糖转移酶 1 通过稳定非特异性致死蛋白 NSL3 来调节整体组蛋白 H4 乙酰化
DOI：
10.1074/jbc.m117.781401
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发表时间：
2017
影响因子：
4.8
通讯作者：
Jin Jingji
中科院分区：
文献类型：
--
作者：
Wu Donglu;Zhao Linhong;Feng Zhitong;Yu Chao;Ding Jian;Wang Lingyao;Wang Fei;Liu Da;Zhu Huihui;Xing Feiyang;Conaway Joan W.;Conaway Ronald C.;Cai Yong;Jin Jingji
人类男性在第一（MOF）含组蛋白乙酰转移酶非特异性致死（NSL）复合体上缺失9个亚基，包括o -连接的n -乙酰氨基葡萄糖（O-GlcNAc）转移酶异构体1 （OGT1）。然而，OGT1的O-GlcNAc转移酶活性是否控制NSL复合物的组蛋白乙酰转移酶活性以及OGT1是否与NSL复合物的其他亚基发生物理相互作用尚不清楚。本研究表明，OGT1主要通过o - glcn酰化组蛋白H4 Lys-16、Lys-5和Lys-8来调节NSL复合物的活性，并稳定NSL复合物亚基NSL3。在人细胞中敲低或过表达OGT1显著影响组蛋白H4残基Lys-16、Lys-5和Lys-8的整体乙酰化。由于OGT1是NSL复合体的一个亚基，我们也研究了OGT1在该复合体中的功能。共转染/共免疫沉淀实验结合体外O-GlcNAc转移酶测定证实，OGT1特异性结合并使O-GlcNAc酰化NSL3。此外，小麦胚芽凝集素亲和纯化证实了细胞中NSL3上存在O-GlcNAc修饰。此外，野生型OGT1 （OGT1- wt）对NSL3的o - glcn酰化作用稳定了NSL3。这种稳定性在NSL3与OGT1突变体OGT1C964A共转染后失去，OGT1突变体OGT1C964A缺乏O-GlcNAc转移酶活性。此外，OGT1-WT对NSL3的稳定显著提高了细胞中H4 - Lys-5、Lys-8和Lys-16的乙酰化水平。这些结果表明，OGT1通过稳定NSL3来调节NSL复合物的活性。
The human males absent on the first (MOF)-containing histone acetyltransferase nonspecific lethal (NSL) complex comprises nine subunits including the O-linked N-acetylglucosamine (O-GlcNAc) transferase, isoform 1 (OGT1). However, whether the O-GlcNAc transferase activity of OGT1 controls histone acetyltransferase activity of the NSL complex and whether OGT1 physically interacts with the other NSL complex subunits remain unclear. Here, we demonstrate that OGT1 regulates the activity of the NSL complex by mainly acetylating histone H4 Lys-16, Lys-5, and Lys-8 via O-GlcNAcylation and stabilization of the NSL complex subunit NSL3. Knocking down or overexpressing OGT1 in human cells remarkably affected the global acetylation of histone H4 residues Lys-16, Lys-5, and Lys-8. Because OGT1 is a subunit of the NSL complex, we also investigated the function of OGT1 in this complex. Co-transfection/co-immunoprecipitation experiments combined with in vitro O-GlcNAc transferase assays confirmed that OGT1 specifically binds to and O-GlcNAcylates NSL3. In addition, wheat germ agglutinin affinity purification verified the occurrence of O-GlcNAc modification on NSL3 in cells. Moreover, O-GlcNAcylation of NSL3 by wild-type OGT1 (OGT1-WT) stabilized NSL3. This stabilization was lost after co-transfection of NSL3 with an OGT1 mutant, OGT1C964A, that lacks O-GlcNAc transferase activity. Furthermore, stabilization of NSL3 by OGT1-WT significantly increased the global acetylation levels of H4 Lys-5, Lys-8, and Lys-16 in cells. These results suggest that OGT1 regulates the activity of the NSL complex by stabilizing NSL3.