Fuller S D, Gowen B E, Reinsch S, Sawyer A, Buendia B, Wepf R, Karsenti E
Structural Biology Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Curr Biol. 1995 Dec 1;5(12):1384-93. doi: 10.1016/s0960-9822(95)00276-4.
The microtubule network, upon which transport occurs in higher cells, is formed by the polymerization of alpha and beta tubulin. The third major tubulin isoform, gamma tubulin, is believed to serve a role in organizing this network by nucleating microtubule growth on microtubule-organizing centers, such as the centrosome. Research in vitro has shown that gamma tubulin must be restored to stripped centrioles to regenerate the centrosomal functions of duplication and microtubule nucleation.
We have re-examined the localization of gamma tubulin in isolated and in situ mammalian centrosomes using a novel immunocytochemical technique that preserves antigenicity and morphology while allowing increased accessibility. As expected, alpha tubulin was localized in cytoplasmic and centriolar barrel microtubules and in the associated pericentriolar material. Foci of gamma tubulin were observed at the periphery of the organized pericentriolar material, as reported previously, often near the termini of microtubules. A further and major location of gamma tubulin was a structure within the proximal end of the centriolar barrel. The distributions were complementary, in that alpha tubulin was excluded from the core of the centriole, and gamma tubulin was excluded from the microtubule barrel.
We have shown that gamma tubulin is localized both in the pericentriolar material and in the core of the mammalian centriole. This result suggests that gamma tubulin has a role in the centriolar duplication process, perhaps as a template for growth of the centriolar microtubules, in addition to its established role in the nucleation of astral microtubules.
在高等细胞中,运输过程所依赖的微管网络是由α和β微管蛋白聚合形成的。微管蛋白的第三种主要异构体γ微管蛋白,被认为通过在诸如中心体等微管组织中心使微管生长成核,从而在组织这个网络中发挥作用。体外研究表明,γ微管蛋白必须被恢复到去除微管蛋白的中心粒上,才能使中心体恢复复制和微管成核的功能。
我们使用一种新颖的免疫细胞化学技术,重新检查了分离的和原位的哺乳动物中心体中γ微管蛋白的定位。该技术在保持抗原性和形态的同时增加了可及性。正如预期的那样,α微管蛋白定位于细胞质和中心粒筒状微管以及相关的中心粒周围物质中。如先前报道,在有组织的中心粒周围物质的周边观察到γ微管蛋白的聚集区,通常靠近微管的末端。γ微管蛋白的另一个主要定位是在中心粒筒近端的一个结构内。这些分布是互补的,因为α微管蛋白被排除在中心粒的核心之外,而γ微管蛋白被排除在微管筒之外。
我们已经表明,γ微管蛋白定位于哺乳动物中心粒周围物质和中心粒的核心中。这一结果表明,γ微管蛋白除了在星体微管成核中已确立的作用外,在中心粒复制过程中也发挥作用,也许作为中心粒微管生长的模板。