Regulation Dynamics of Leishmania Differentiation: Deconvoluting Signals and Identifying Phosphorylation Trends*
Regulation Dynamics of Leishmania Differentiation: Deconvoluting Signals and Identifying Phosphorylation Trends*
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利什曼原虫分化的调控动态：解卷积信号和识别磷酸化趋势*
DOI：
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发表时间：
2014
影响因子：
7
通讯作者：
D. Zilberstein
中科院分区：
文献类型：
--
作者：
P. Tsigankov;P. Gherardini;M. Helmer;G. Späth;P. Myler;D. Zilberstein
利什曼原虫是一种必需的细胞内寄生原虫，可引起人类多种疾病，在沙蝇中肠的胞外前鞭毛体和哺乳动物巨噬细胞吞噬溶酶体的胞内无鞭毛体之间循环。虽然巨噬细胞内发育的许多分子机制仍然是一个谜，但模拟吞噬溶酶体条件(37℃和pH 5.5)的无宿主系统的发展为这些过程提供了新的见解。前鞭毛体向无鞭毛体分化的过程在形态上可分为4个不同的阶段：I，信号感受(暴露后0~5h)，II，运动停止和聚集(5~10h)，III，无鞭毛体形态发生(10~24h)，IV，成熟(24~120h)。转录学和蛋白质组学分析表明，分化是一个协调的过程，导致对吞噬溶酶体内的生活的适应。最近的磷酸蛋白质组学分析表明，前鞭毛体和无鞭毛体之间的磷酸化存在着广泛的差异，并确定了阶段特异性的磷酸化基序。我们假设分化信号激活了启动利什曼原虫转化的磷酸化途径，在这里，我们使用等压标记进行相对和绝对定量，以询问在杜氏利什曼原虫前鞭毛体到无鞭毛体分化过程中磷酸化谱变化的动态。对来自106个蛋白质的163个磷酸肽的分析显示了6个不同的动力学特征；在I和III阶段以磷酸化增加为主，而II和IV阶段以更大的去磷酸化为特征。几种蛋白质(包括蛋白激酶)在暴露于完全分化信号(即，信号特异性；37℃和pH 5.5)后在I相被磷酸化，但在两个物理参数中的任何一个单独之后都不被磷酸化。其他几种蛋白激酶(包括调节亚基)和磷酸酶在分化过程中也显示出磷酸化的变化。这项工作构成了对寄生原生动物中磷酸化动力学的首次基因组规模的询问，揭示了利什曼原虫分化过程中的信号通路的轮廓。质谱学蛋白质组学数据已保存到ProteomeXchange联盟(识别符PXD000671)。可以使用ProteinPilot™软件查看数据。
Leishmania are obligatory intracellular parasitic protozoa that cause a wide range of diseases in humans, cycling between extracellular promastigotes in the mid-gut of sand flies and intracellular amastigotes in the phagolysosomes of mammalian macrophages. Although many of the molecular mechanisms of development inside macrophages remain a mystery, the development of a host-free system that simulates phagolysosome conditions (37 °C and pH 5.5) has provided new insights into these processes. The time course of promastigote-to-amastigote differentiation can be divided into four morphologically distinct phases: I, signal perception (0–5 h after exposure); II, movement cessation and aggregation (5–10 h); III, amastigote morphogenesis (10–24 h); and IV, maturation (24–120 h). Transcriptomic and proteomic analyses have indicated that differentiation is a coordinated process that results in adaptation to life inside phagolysosomes. Recent phosphoproteomic analysis revealed extensive differences in phosphorylation between promastigotes and amastigotes and identified stage-specific phosphorylation motifs. We hypothesized that the differentiation signal activates a phosphorylation pathway that initiates Leishmania transformation, and here we used isobaric tags for relative and absolute quantitation to interrogate the dynamics of changes in the phosphorylation profile during Leishmania donovani promastigote-to-amastigote differentiation. Analysis of 163 phosphopeptides (from 106 proteins) revealed six distinct kinetic profiles; with increases in phosphorylation predominated during phases I and III, whereas phases II and IV were characterized by greater dephosphorylation. Several proteins (including a protein kinase) were phosphorylated in phase I after exposure to the complete differentiation signal (i.e. signal-specific; 37 °C and pH 5.5), but not after either of the physical parameters separately. Several other protein kinases (including regulatory subunits) and phosphatases also showed changes in phosphorylation during differentiation. This work constitutes the first genome-scale interrogation of phosphorylation dynamics in a parasitic protozoa, revealing the outline of a signaling pathway during Leishmania differentiation. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (identifier PXD000671). Data can be viewed using ProteinPilot™ software.
影响因子：
4
作者：
Debrabant, A;Joshi, MB;Dwyer, DM
通讯作者：
Dwyer, DM
影响因子：
4.4
作者：
Liao, Lujian;McClatchy, Daniel B.;Park, Sung Kyu;Xu, Tao;Lu, Bingwen;Yates, John R., III
通讯作者：
Yates, John R., III