O-GlcNAc-Modification of NSL3 at Thr755 Site Maintains the Holoenzyme Activity of MOF/NSL Histone Acetyltransferase Complex
O-GlcNAc-Modification of NSL3 at Thr755 Site Maintains the Holoenzyme Activity of MOF/NSL Histone Acetyltransferase Complex
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NSL3 Thr755 位点的 O-GlcNAc 修饰维持 MOF/NSL 组蛋白乙酰转移酶复合物的全酶活性
DOI：
10.3390/ijms21010173
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发表时间：
2019-12
影响因子：
5.6
通讯作者：
Jingji Jin
中科院分区：
文献类型：
--
作者：
Linhong Zhao;Min Li;Tao Wei;Chang Feng;Tingting Wu;Junaid Ali Shah;Hongsen Liu;Fei Wang;Yong Cai;Jingji Jin
O-连锁β-N-乙酰氨基葡萄糖(O-GlcNAc)转移酶亚型1(OGT1)和非特异性致死蛋白3(NSL3)都是MOF/NSL组蛋白-乙酰基转移酶复合体的重要组成部分。我们先前描述了组蛋白H4乙酰化水平是如何受OGT1/O-GlcNacylation介导的NSL3稳定性调节的。然而，NSL3的特异性修饰位点及其影响蛋白质稳定性的分子机制尚不清楚。在这里，我们提出了来自生化实验的证据，证明NSL3在Thr755处的O-GlcN酰化与MOF/NSL复合体的全酶活性密切相关。利用体外O-GlcNAc转移酶分析结合质谱学分析，我们推测NSL3 C末端的Thr755残基是由OGT1修饰的O-GlcNAc。重要的是，该位点的O-GlcN酰化参与了NSL3泛素降解的调节，因为该位点突变(T755A)促进了泛素介导的NSL3的降解。进一步的深入研究发现，泛素结合酶E_2·S(UBE_2S)通过直接与NSL3结合，加速了NSL3的降解。有趣的是，OGT1和UBE2S竞争性地与NSL3结合，提示OGT1-UBE2S在调节NSL3稳定性方面具有协同作用。此外，NSL3 Thr755位点的O-GlcN酰化调节了组蛋白H4在赖氨酸5、8和16处的乙酰化水平，这表明NSL3在Thr755处的O-GlcN酰化是维持MOF/NSL复合体的完整性和全酶活性所必需的。在集落形成试验中，我们发现复合体的完整性影响肺腺癌II型上皮样A549细胞的增殖。综上所述，我们的结果为阐明MOF/NSL复合体的分子机制提供了新的视角。
Both OGT1 (O-linked β-N-acetylglucosamine (O-GlcNAc) transferase isoform 1) and NSL3.(nonspecific lethal protein 3) are crucial components of the MOF (males absent on the first)/NSL histone.acetyltransferase complex. We previously described how global histone H4 acetylation levels were.modulated by OGT1/O-GlcNAcylation-mediated NSL3 stability. However, the specific modification.site of NSL3 and its molecular mechanism of protein stability remain unknown. Here, we present.evidence from biochemical experiments arguing that O-GlcNAcylation of NSL3 at Thr755 is tightly.associated with holoenzyme activity of the MOF/NSL complex. Using in vitro O-GlcNAc-transferase.assays combined with mass spectrometry, we suppose that the residue Thr755 on NSL3 C-terminus is.the major site O-GlcNAc-modified by OGT1. Importantly, O-GlcNAcylation of this site is involved in.the regulation of the ubiquitin-degradation of NSL3, because this site mutation (T755A) promotes the.ubiquitin-mediated degradation of NSL3. Further in-depth research found that ubiquitin conjugating.enzyme E2 S (UBE2S) accelerated the degradation of NSL3 via direct binding to it. Interestingly,.OGT1 and UBE2S competitively bind to NSL3, suggesting the coordination of OGT1–UBE2S in.regulating NSL3 stability. Furthermore, O-GlcNAcylation of NSL3 Thr755 site regulates the histone.H4 acetylation levels at lysine 5, 8, and 16, suggesting that the O-GlcNAcylation of NSL3 at Thr755 is.required for maintaining the integrity and holoenzyme activity of the MOF/NSL complex. In colony.formation assays, we found that the integrity of the complex impacts the proliferation of the lung.carcinoma type II epithelium-like A549 cells. Taken together, our results provide new insight into the.elucidation of the molecular mechanism of the MOF/NSL complex.
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影响因子：
29
作者：
Ruan HB;Han X;Li MD;Singh JP;Qian K;Azarhoush S;Zhao L;Bennett AM;Samuel VT;Wu J;Yates JR 3rd;Yang X
通讯作者：
Yang X
影响因子：
8
作者：
通讯作者：
--
影响因子：
3.5
作者：
L. Wells;G. Hart
通讯作者：
L. Wells;G. Hart
影响因子：
4
作者：
Geng, Fei;Zhu, Weijia;Andrews, David W.
通讯作者：
Andrews, David W.
影响因子：
5.7
作者：
Liess, Anna K. L.;Kucerova, Alena;Lorenz, Sonja
通讯作者：
Lorenz, Sonja