Analysis of dynamic changes in post-translational modifications of human histones during cell cycle by mass spectrometry
Analysis of dynamic changes in post-translational modifications of human histones during cell cycle by mass spectrometry
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DOI：
10.1074/mcp.m700070-mcp200
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发表时间：
2007-11-01
影响因子：
7
通讯作者：
van Oostrum, Jan
中科院分区：
文献类型：
--
作者：
Bonenfant, Debora;Towbin, Harry;van Oostrum, Jan
四个核心组蛋白的N-末端尾部经历几种类型的共价翻译后修饰，其在调节染色质结构和功能中具有特定作用。在这里，我们提出了一个广泛的分析发生在细胞周期的核心组蛋白修饰。我们的MS实验表征了G1、S和G（2）/M期阻滞的HeLa细胞组蛋白的修饰模式。对于所有的核心组蛋白，在G1和S期的修改基本相同，但在有丝分裂过程中有很大的不同。使用SILAC（细胞培养物中氨基酸的稳定同位素标记）方法定量S和G（2）/M期之间的修饰变化。最引人注目的是组蛋白H3和H4上的有丝分裂磷酸化，而H2 A上的磷酸化在细胞周期中是恒定的。在G2/M期阻滞的细胞中，观察到所有组蛋白的乙酰化丧失。周期依赖性甲基化的模式更为复杂：在G（2）/M期间，H3 Lys（27）和Lys（36）减少，而H4 Lys（20）增加。我们的研究结果表明，有丝分裂是细胞周期的一个时期，在此期间，许多修饰表现出动态变化。
The N-terminal tails of the four core histones are subject to several types of covalent post-translational modifications that have specific roles in regulating chromatin structure and function. Here we present an extensive analysis of the core histone modifications occurring through the cell cycle. Our MS experiments characterized the modification patterns of histones from HeLa cells arrested in phase G1, S, and G(2)/M. For all core histones, the modifications in the G1 and S phases were largely identical but drastically different during mitosis. Modification changes between S and G(2)/M phases were quantified using the SILAC (stable isotope labeling by amino acids in cell culture) approach. Most striking was the mitotic phosphorylation on histone H3 and H4, whereas phosphorylation on H2A was constant during the cell cycle. A loss of acetylation was observed on all histones in G2/M-arrested cells. The pattern of cycle-dependent methylation was more complex: during G(2)/M, H3 Lys(27) and Lys(36) were decreased, whereas H4 Lys(20) was increased. Our results show that mitosis was the period of the cell cycle during which many modifications exhibit dynamic changes.