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Mitogen-Activated Protein Kinase Kinase 5 Regulates Proliferation and Biosynthetic Processes in Procyclic Forms of Trypanosoma brucei
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2017-11-07 00:00:00 , DOI: 10.1021/acs.jproteome.7b00415
Fernanda G. Kugeratski 1, 2 , Michel Batista , Carla V. de Paula Lima , Lisa J. Neilson 1 , Elizabeth Sousa da Cunha , Lyris M. de Godoy , Sara Zanivan 1, 2 , Marco A. Krieger , Fabricio K. Marchini
Affiliation  

The pathogenic protozoan T. brucei alternates into distinct developmental stages in the mammalian and insect hosts. The mitogen-activated protein kinase (MAPK) signaling pathways transduce extracellular stimuli into a range of cellular responses, which ultimately lead to the adaptation to the external environment. Here, we combined a loss of function approach with stable isotope labeling with amino acids in cell culture (SILAC)-based mass spectrometry (MS) to investigate the role of the mitogen-activated protein kinase kinase 5 (MKK5) in T. brucei. The silencing of MKK5 significantly decreased the proliferation of procyclic forms of T. brucei. To shed light on the molecular alterations associated with this phenotype, we measured the total proteome and phosphoproteome of cells silenced for MKK5. In the total proteome, we observed a general decrease in proteins related to ribosome and translation as well as down-regulation of several components of the fatty acids biosynthesis pathway. In addition, we observed alterations in the protein levels and phosphorylation of key metabolic enzymes, which point toward a suppression of the oxidative metabolism. Taken together, our findings show that the silencing of MKK5 alters cell growth, energy metabolism, protein and fatty acids biosynthesis in procyclic T. brucei.

中文翻译:

丝裂原激活的蛋白激酶激酶5调节布鲁氏锥虫的前环形式的增殖和生物合成过程。

致病性原生动物布鲁氏菌在哺乳动物和昆虫宿主中交替进入不同的发育阶段。丝裂原激活的蛋白激酶(MAPK)信号通路将细胞外刺激转化为一系列细胞反应,最终导致对外部环境的适应。在这里,我们在组合的函数逼近与稳定同位素标记损失与在细胞培养物(SILAC)基质谱(MS)的氨基酸来研究促分裂原活化蛋白激酶的作用激酶5(MKK5)布氏锥虫。MKK5的沉默显著下降的procyclic形式增殖布氏锥虫。为了阐明与该表型有关的分子变化,我们测量了沉默了MKK5的细胞的总蛋白质组和磷酸化蛋白质组。在总蛋白质组中,我们观察到与核糖体和翻译有关的蛋白质普遍减少,以及脂肪酸生物合成途径的某些成分的下调。此外,我们观察到蛋白质水平和关键代谢酶磷酸化的改变,这表明氧化代谢受到抑制。总之,我们的研究结果表明,MKK5会改变细胞的生长,能量代谢,蛋白质和脂肪酸合成中procyclic的沉默布氏锥虫
更新日期:2017-11-08
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