Global turnover of histone post-translational modifications and variants in human cells.
Global turnover of histone post-translational modifications and variants in human cells.
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DOI：
10.1186/1756-8935-3-22
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发表时间：
2010-12-06
影响因子：
3.9
通讯作者：
Garcia BA
中科院分区：
文献类型：
--
作者：
Zee BM;Levin RS;DiMaggio PA;Garcia BA
组蛋白和组蛋白变体的N-末端尾部上的翻译后修饰（PTM）调节不同的转录状态和核事件。而特定的PTM的功能效应是目前的主题，激烈的调查，大多数研究组蛋白PTM/变体的非时间的方式，很少有研究报道这些组蛋白形式的动力学信息。以前的研究已经使用放射性标记，荧光显微镜和染色质免疫沉淀来确定组蛋白的周转率，并发现了有趣的周转增加和基因表达增加之间的相关性。因此，组蛋白周转是一个研究不足，但潜在的重要参数，可能有助于表观遗传调控。理解组蛋白修饰和序列变异背景下的营业额可以为组蛋白替换功能提供有价值的额外见解。在这项研究中，我们通过将细胞培养中氨基酸的稳定同位素标记（SILAC）脉冲实验与基于定量质谱的蛋白质组学相结合，测量了标记同位素掺入HeLa细胞组蛋白的代谢率。总的来说，我们发现大多数核心组蛋白具有相似的周转率，除了H2 A变体，其表现出更广泛的速率，可能与其表观遗传功能一致。此外，乙酰化的组蛋白有一个显着更快的营业额相比，一般的组蛋白和甲基化的组蛋白，虽然这些速率变化很大，这取决于网站和整体甲基化程度。一直发现含有转录活性标记的组蛋白比含有沉默标记的组蛋白具有更快的周转率。有趣的是，在同一肽上同时存在活性标记和沉默标记导致比在同一肽上单独存在任一标记更慢的周转率。最后，我们观察到H3.1、H3.2和H3.3变体的几乎所有修饰形式之间的周转率几乎没有差异，值得注意的例外是H3.2K36me2在其他H3变体上的周转率比该标记更快。定量蛋白质组学提供了互补的洞察力，以前的工作，旨在定量测量组蛋白营业额，我们的研究结果表明，营业额取决于特定位点的翻译后修饰和序列变异。
Post-translational modifications (PTMs) on the N-terminal tails of histones and histone variants regulate distinct transcriptional states and nuclear events. Whereas the functional effects of specific PTMs are the current subject of intense investigation, most studies characterize histone PTMs/variants in a non-temporal fashion and very few studies have reported kinetic information about these histone forms. Previous studies have used radiolabeling, fluorescence microscopy and chromatin immunoprecipitation to determine rates of histone turnover, and have found interesting correlations between increased turnover and increased gene expression. Therefore, histone turnover is an understudied yet potentially important parameter that may contribute to epigenetic regulation. Understanding turnover in the context of histone modifications and sequence variants could provide valuable additional insight into the function of histone replacement. In this study, we measured the metabolic rate of labeled isotope incorporation into the histone proteins of HeLa cells by combining stable isotope labeling of amino acids in cell culture (SILAC) pulse experiments with quantitative mass spectrometry-based proteomics. In general, we found that most core histones have similar turnover rates, with the exception of the H2A variants, which exhibit a wider range of rates, potentially consistent with their epigenetic function. In addition, acetylated histones have a significantly faster turnover compared with general histone protein and methylated histones, although these rates vary considerably, depending on the site and overall degree of methylation. Histones containing transcriptionally active marks have been consistently found to have faster turnover rates than histones containing silent marks. Interestingly, the presence of both active and silent marks on the same peptide resulted in a slower turnover rate than either mark alone on that same peptide. Lastly, we observed little difference in the turnover between nearly all modified forms of the H3.1, H3.2 and H3.3 variants, with the notable exception that H3.2K36me2 has a faster turnover than this mark on the other H3 variants. Quantitative proteomics provides complementary insight to previous work aimed at quantitatively measuring histone turnover, and our results suggest that turnover rates are dependent upon site-specific post-translational modifications and sequence variants.
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影响因子：
3.7
作者：
Clausell J;Happel N;Hale TK;Doenecke D;Beato M
通讯作者：
Beato M
影响因子：
64.8
作者：
Fischle, W;Wang, YM;Allis, CD
通讯作者：
Allis, CD
影响因子：
56.9
作者：
Dion, Michael F.;Kaplan, Tommy;Rando, Oliver J.
通讯作者：
Rando, Oliver J.
DOI：
10.1073/pnas.0600803103
发表时间：
2006-04-25
影响因子：
11.1
作者：
Hake, SB;Allis, CD
通讯作者：
Allis, CD
DOI：
10.1073/pnas.0507975102
发表时间：
2005-12-20
影响因子：
11.1
作者：
Li, B;Pattenden, SG;Workman, JL
通讯作者：
Workman, JL