Brodsky I B, Burakov A V, Nadezhdina E S
Group of Cell Biology, Institute of Protein Research of Russian Academy of Science, Moscow, Russia.
Cell Motil Cytoskeleton. 2007 Jun;64(6):407-17. doi: 10.1002/cm.20192.
Microtubules in interphase mammalian cells usually form a radial array with minus-ends concentrated in the central region and plus-ends placed at the periphery. This is accepted as correct, that two factors determinate the radial organization of microtubules - the centrosome, which nucleate and anchor the microtubules minus-ends, and the interaction of microtubules with cortical dynein, which positions centrosome in the cell center. However, it looks as if there are additional factors, affecting the radial structure of microtubule system. We show here that in aged Vero cytoplasts (17 h after enucleation) microtubule system lost radial organization and became chaotic. To clear up the reasons of that, we studied centrosome activity, its position in the cytoplasts and microtubule dynamics. We found that centrosome in aged cytoplasts was still active and placed in the central region of the cytoplasm, while after total disruption of the microtubules it was displaced from the center. Microtubules in aged cytoplasts were not stabilized, but they lost their ability to stop to grow near cell cortex and continued to grow reaching it. Aged cytoplast lamellae was partially depleted with dynactin though Golgi remained compact indicating dynein activity. We conclude that microtubule stoppage at cell cortex is mediated by some (protein) factors, and these factors influence radial structure of microtubule system. It seems that the key role in centrosome positioning is played by dynein complexes anchored everywhere in the cytoplasm rather than anchored in cell cortex.
处于间期的哺乳动物细胞中的微管通常形成一种放射状排列,负端集中在中央区域,正端位于周边。人们认为以下两个因素决定了微管的放射状排列,这一点是正确的:中心体,它使微管负端成核并锚定;以及微管与皮层动力蛋白的相互作用,它将中心体定位在细胞中心。然而,似乎还有其他因素影响微管系统的放射状结构。我们在此表明,在老化的非洲绿猴肾细胞胞质体(去核后17小时)中,微管系统失去了放射状排列并变得混乱。为了弄清楚其中的原因,我们研究了中心体活性、它在胞质体中的位置以及微管动力学。我们发现老化胞质体中的中心体仍然活跃并位于细胞质的中央区域,而在微管完全破坏后它从中心移位。老化胞质体中的微管没有得到稳定,但它们失去了在细胞皮层附近停止生长的能力,继续生长并到达皮层。老化胞质体的板层中动力蛋白激活蛋白部分耗尽,尽管高尔基体仍然紧密,这表明动力蛋白有活性。我们得出结论,微管在细胞皮层的停止生长是由一些(蛋白质)因子介导的,并且这些因子影响微管系统的放射状结构。似乎在中心体定位中起关键作用的是锚定在细胞质各处而非锚定在细胞皮层的动力蛋白复合物。