In Vivo Residue-specific Histone Methylation Dynamics
In Vivo Residue-specific Histone Methylation Dynamics
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DOI：
10.1074/jbc.m109.063784
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发表时间：
2010-01-29
影响因子：
4.8
通讯作者：
Garcia, Benjamin A.
中科院分区：
文献类型：
--
作者：
Zee, Barry M.;Levin, Rebecca S.;Garcia, Benjamin A.
特定组蛋白残基的甲基化能够激活基因，也能使基因沉默。尽管在甲基化功能方面进行了大量研究，但大多数研究要么呈现的是甲基化的静态快照，要么无法将动力学信息赋予特定残基。我们利用高分辨率质谱仪上的液相色谱 - 串联质谱法以及重甲基 - 稳定同位素标记氨基酸细胞培养（heavy methyl - SILAC）标记，研究了位点特异性组蛋白赖氨酸和精氨酸的甲基化动态。对处于一种甲基化状态的标记中间产物的检测表明，单甲基化、双甲基化和三甲基化残基的形成速率通常逐渐变慢。此外，与活跃基因相关的甲基化比与沉默基因相关的甲基化速率更快。最后，在同一肽段上同时存在激活标记和沉默标记时，其甲基化速率比单独存在任何一种标记时都要慢。在此我们表明，像这样的定量蛋白质组学方法能够确定多个甲基化残基的动态，这是组蛋白生物学中一个研究不足的部分。
Methylation of specific histone residues is capable of both gene activation and silencing. Despite vast work on the function of methylation, most studies either present a static snapshot of methylation or fail to assign kinetic information to specific residues. Using liquid chromatography-tandem mass spectrometry on a high-resolution mass spectrometer and heavy methyl-SILAC labeling, we studied site-specific histone lysine and arginine methylation dynamics. The detection of labeled intermediates within a methylation state revealed that mono-, di-, and trimethylated residues generally have progressively slower rates of formation. Furthermore, methylations associated with active genes have faster rates than methylations associated with silent genes. Finally, the presence of both an active and silencing mark on the same peptide results in a slower rate of methylation than the presence of either mark alone. Here we show that quantitative proteomic approaches such as this can determine the dynamics of multiple methylated residues, an understudied portion of histone biology.