Middle-Down Characterization of the Cell Cycle Dependence of Histone H4 Posttranslational Modifications and Proteoforms.
Middle-Down Characterization of the Cell Cycle Dependence of Histone H4 Posttranslational Modifications and Proteoforms.
复制标题
DOI：
10.1002/pmic.201700442
复制
发表时间：
2018-06
期刊：
影响因子：
3.4
通讯作者：
Young NL
中科院分区：
文献类型：
--
作者：
Jiang T;Hoover ME;Holt MV;Freitas MA;Marshall AG;Young NL
组蛋白的翻译后修饰（PTM）是重要的表观遗传调节机制，其在癌症中经常失调。我们采用中间蛋白质组学研究组蛋白H4在细胞周期进程中的PTM和蛋白质型。采用pH梯度弱阳离子交换-亲水作用液相色谱（WCX-HILIC）进行在线液相色谱-质谱分析，对组蛋白H4的蛋白质组进行分离和分析。该方法提供了蛋白质形式（包括位置异构体）的增强分离，并简化了下游数据分析。我们使用高质量精度和分辨率的傅立叶变换离子回旋共振（FT-ICR）质谱仪来明确区分乙酰化和三甲基化（Δ m = 0.036 Da）。总之，我们确定和量化233组蛋白H4在两个乳腺癌细胞系中的蛋白形式。我们观察到有丝分裂过程中S1磷酸化的显着增加，暗示有丝分裂染色质凝聚的重要作用。随着细胞周期的进展，观察到K20未修饰的蛋白形式减少，对应于K20单甲基化和二甲基化的增加。随着细胞从S期进入有丝分裂，K5、K8、K12和K16的乙酰化水平下降，表明细胞周期依赖性，在染色质复制和浓缩过程中发挥重要作用。这些对细胞周期表观遗传学的新见解可能提供新的诊断和预后生物标志物。
Post-translational modifications (PTMs) of histones are important epigenetic regulatory mechanisms that are often dysregulated in cancer. We employ middle-down proteomics to investigate the PTMs and proteoforms of histone H4 during cell cycle progression. We use pH gradient weak cation exchange-hydrophilic interaction liquid chromatography (WCX-HILIC) for on-line liquid chromatography-mass spectrometry analysis to separate and analyze the proteoforms of histone H4. This procedure provides enhanced separation of proteoforms, including positional isomers, and simplifies downstream data analysis. We use ultrahigh mass accuracy and resolution Fourier transform-ion cyclotron resonance (FT-ICR) mass spectrometer to unambiguously distinguish between acetylation and tri-methylation (Δm = 0.036 Da). In total, we identify and quantify 233 proteoforms of histone H4 in two breast cancer cell lines. We observe significant increases in S1 phosphorylation during mitosis, implicating an important role in mitotic chromatin condensation. A decrease of K20 unmodified proteoforms is observed as the cell cycle progresses, corresponding to an increase of K20 mono- and di-methylation. Acetylation at K5, K8, K12, and K16 declines as cells traverse from S phase to mitosis, suggesting cell cycle–dependence and an important role during chromatin replication and condensation. These new insights into the epigenetics of the cell cycle may provide new diagnostic and prognostic biomarkers.
登录
查看更多内容
影响因子：
2.1
作者：
Lo PK;Sukumar S
通讯作者：
Sukumar S
影响因子：
1.6
作者：
Barber, CM;Turner, FB;Allis, CD
通讯作者：
Allis, CD
影响因子：
9.2
作者：
Fuchs, Stephen M.;Krajewski, Krzysztof;Baker, Richard W.;Miller, Victoria L.;Strahl, Brian D.
通讯作者：
Strahl, Brian D.
影响因子：
64.8
作者：
Moriniere, Jeanne;Rousseaux, Sophie;Petosa, Carlo
通讯作者：
Petosa, Carlo
影响因子：
7
作者：
Chen, Yu;Hoover, Michael E.;Young, Nicolas L.
通讯作者：
Young, Nicolas L.