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ADP核糖基化因子样蛋白2(Arl2)影响乳腺癌细胞中的微管动力学。

ADP ribosylation factor like 2 (Arl2) protein influences microtubule dynamics in breast cancer cells.

作者信息

Beghin Anne, Honore Stephane, Messana Celine, Matera Eva-Laure, Aim Jennifer, Burlinchon Sandrine, Braguer Diane, Dumontet Charles

机构信息

Laboratoire de Cytologie Analytique, UMR INSERM 590; Université de Lyon FR69008, Lyon, France.

出版信息

Exp Cell Res. 2007 Feb 1;313(3):473-85. doi: 10.1016/j.yexcr.2006.10.024. Epub 2006 Oct 28.

Abstract

ADP ribosylation factor like 2 (Arl2) protein is involved in the folding of tubulin peptides. Variants of the human adenocarcinoma line MCF7 cells with increased or reduced content of Arl2 protein were produced and characterized. Western blot analysis performed after separation of the different fractions of tubulins showed that the content in polymerizable soluble heterodimers was significantly increased in cells with the highest Arl2 expression level (MA+) and reduced in cells with the lowest Arl2 expression level (MA-) in comparison to control cells (MP). Microtubule dynamic instability, measured after microinjection of rhodamine-labelled tubulin in living cells, was significantly enhanced in MA+ cells and reduced in MA- cells. These alterations involved modifications of the microtubule growth and shortening rates, duration of attenuation phases, percentage of time spent in each phase (growth, shortening and attenuation) and catastrophe frequency. We also observed modifications in the expression level of the tumor suppressor protein phosphatase 2Ac, which has been shown to form a complex with Arl2. Finally, cell cycle progression was modified in these cells, particularly in regard to duration of telophase. In summary, alterations in Arl2 protein content were found to be associated with modifications in tubulin pools, microtubule dynamics as well as cell cycle progression.

摘要

ADP核糖基化因子样蛋白2(Arl2)参与微管蛋白肽的折叠。制备并表征了人腺癌系MCF7细胞中Arl2蛋白含量增加或减少的变体。在分离不同微管蛋白组分后进行的蛋白质印迹分析表明,与对照细胞(MP)相比,Arl2表达水平最高的细胞(MA +)中可聚合可溶性异二聚体的含量显著增加,而Arl2表达水平最低的细胞(MA-)中可聚合可溶性异二聚体的含量则降低。在活细胞中显微注射罗丹明标记的微管蛋白后测量的微管动态不稳定性,在MA +细胞中显著增强,而在MA-细胞中降低。这些改变涉及微管生长和缩短速率、衰减期持续时间、每个阶段(生长、缩短和衰减)所花费时间的百分比以及灾难频率的改变。我们还观察到肿瘤抑制蛋白磷酸酶2Ac的表达水平发生了改变,该蛋白已被证明与Arl2形成复合物。最后,这些细胞的细胞周期进程发生了改变,特别是在末期持续时间方面。总之,发现Arl2蛋白含量的改变与微管蛋白库、微管动力学以及细胞周期进程的改变有关。

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