Confocal investigation on colocalization between tubulin posttranslational modifications and associated proteins in rat C6 glioma cells
Confocal investigation on colocalization between tubulin posttranslational modifications and associated proteins in rat C6 glioma cells
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DOI：
10.1016/j.jsb.2020.107676
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发表时间：
2021-03-01
影响因子：
3
通讯作者：
Gadau, Sergio D.
中科院分区：
文献类型：
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作者：
Arru, Caterina;Serra, Elisa;Gadau, Sergio D.
多形性胶质母细胞瘤是最致命的脑肿瘤。在其发展机制的研究中，人们关注的是其细胞的微管网络，主要是β III微管蛋白，它被认为是恶性肿瘤的标志。在本研究中，我们选择研究胶质母细胞瘤细胞中的微管蛋白编码，分析微管蛋白翻译后修饰与与之相关的不同蛋白之间的相互作用程度。我们评估了EB-1、CLIP-170和kinesin-1等多种相关蛋白的模式，以及它们与翻译后最丰富的微管蛋白修饰（酪氨酸化、乙酰化和多谷氨酰化）的共分布程度。通过免疫荧光，我们发现EB-1， CLIP-170和kinesin-1在胶质母细胞瘤细胞中可以很好地检测到。微管蛋白和相关蛋白翻译后修饰的双荧光和共定位指数显示，酪氨酸化的a-微管蛋白与EB-1、CLIP-170和激酶蛋白-1具有显著的高亲和力，而乙酰化和多谷氨酰化的微管蛋白与这三种相关蛋白的相互作用程度不太明显。本文中提出的数据强调了对胶质母细胞瘤细胞的微管力学进行深入分析的重要性。这可能表明新的实验性治疗方法能够更有选择性地作用于这种类型癌症的细胞微管网络。
Glioblastoma multiforme is the most lethal brain tumor. In the study of mechanisms underlying its development attention has been paid to the microtubular network of its cells, mainly on beta III tubulin, considered as a marker of malignancy. In the present work, we chose to investigate the tubulin code in glioblastoma cells, analyzing the degree of interaction between tubulin post-translational modifications and different proteins associated with them. The pattern of diverse associated proteins such as EB-1, CLIP-170 and kinesin-1 and their degree of co-distribution with the most abundant post-translational tubulin modifications (tyrosination, acetylation and polyglutamylation) were evaluated. Through immunofluorescence we have shown that EB-1, CLIP-170 and kinesin-1 were well detectable in glioblastoma cells. The double fluorescence and colocalization index between the post-translational modifications of tubulin and associated proteins showed that tyrosinated a-tubulin has significantly high affinity with EB-1, CLIP-170 and kinesin-1, while for acetylated and polyglutamylated tubulin, the degree of interaction with the three associated proteins evaluated was less apparent. Data presented in this paper underline the importance of a thorough analysis of the microtubular mechanics in glioblastoma cells. This may suggest new experimental therapeutic approaches able to act more selectively on the microtubular network of cells in this type of cancer.