Instituto de Investigaciones en Ingeniería Genética y Biología Molecular Dr. H.N. Torres, INGEBI-CONICET, Vuelta de Obligado 2490, C1428ADN Buenos Aires, Argentina.
Exp Parasitol. 2012 Dec;132(4):537-45. doi: 10.1016/j.exppara.2012.09.002. Epub 2012 Sep 13.
In eukaryotes, an oscillating network of protein kinase activities drives the order and timing of the cell cycle progression. Complexes formed by cyclins associated to cyclin-dependent kinases (CDKs) are the central components of this network. Cyclins act as the activating subunits and their abundance is regulated by different mechanisms in order to promote or prevent kinase activity. Protein synthesis, proteasomal degradation and/or differential subcellular compartmentalization modulate cyclin expression levels along the cell cycle. We describe in this work the characterization of Trypanosoma cruzi Cyclin 2 (TcCYC2), which contributes to a better understanding of the cell cycle regulation in this protozoan parasite. We found TcCYC2 exhibited cyclin function in a yeast complementation assay and over-expression of hemagglutinin tagged TcCYC2-HA rendered shorter duplication times and smaller cell sizes in the epimastigote form of the parasite. Analysis of synchronized cultures showed that over-expression of TcCYC2-HA altered the timing epimastigotes pass through G2/M boundary or cytokinesis. Taken together, our results showed that TcCYC2 is a functional cyclin whose over-expression modifies the dynamics of the cell cycle as well as the morphology of epimastigote forms of T. cruzi, suggesting it plays an important role in the cell cycle regulation machinery.
在真核生物中,蛋白质激酶活性的振荡网络驱动细胞周期进程的有序性和时间性。由细胞周期蛋白与细胞周期蛋白依赖性激酶 (CDKs) 形成的复合物是该网络的核心组成部分。细胞周期蛋白作为激活亚基,其丰度受到不同机制的调节,以促进或抑制激酶活性。蛋白质合成、蛋白酶体降解和/或差异的细胞区室化沿细胞周期调节细胞周期蛋白的表达水平。我们在这项工作中描述了克氏锥虫细胞周期蛋白 2 (TcCYC2) 的特征,这有助于更好地理解这种原生动物寄生虫的细胞周期调控。我们发现 TcCYC2 在酵母互补测定中表现出细胞周期蛋白功能,并且过表达带有血凝素标签的 TcCYC2-HA 使寄生虫的前鞭毛体形式的复制时间缩短和细胞体积变小。对同步培养物的分析表明,TcCYC2-HA 的过表达改变了 epimastigote 通过 G2/M 边界或胞质分裂的时间。总之,我们的结果表明 TcCYC2 是一种功能性细胞周期蛋白,其过表达改变了细胞周期的动力学以及克氏锥虫前鞭毛体形式的形态,表明它在细胞周期调控机制中发挥着重要作用。