Microtubule dynamics investigated by microinjection of Paramecium axonemal tubulin: lack of nucleation but proximal assembly of microtubules at the kinetochore during prometaphase.
Microtubule dynamics investigated by microinjection of Paramecium axonemal tubulin: lack of nucleation but proximal assembly of microtubules at the kinetochore during prometaphase.
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DOI：
10.1083/jcb.108.3.939
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发表时间：
1989-03
期刊：
影响因子：
--
通讯作者：
DeBrabander M
中科院分区：
文献类型：
--
作者：
Geuens G;Hill AM;Levilliers N;Adoutte A;DeBrabander M
使用未修饰的草履虫纤毛微管蛋白和延时固定在体内研究了PtK2细胞的微管（MT）动力学。利用一种不与脊椎动物细胞质微管蛋白发生反应的特异性抗体定位轴突微管蛋白的结合位点（Adoutte, a .， M. Claisse， R. Maunoury和J. Beisson, 1985）。J. Mol. evolution . 22:220-229)，然后对相同的细胞进行免疫金标记、纳米显微镜和超微结构观察。我们证实了在其他细胞类型中显微注射标记微管蛋白的数据（Soltys, b.j.和g.g. Borisy, 1985）。[j] .中国生物医学工程学报，2010:391 - 391；Mitchison， T.， L. Evans， E. Schulze和M. Kirschner 1986。细胞。45:515 - 527;Schulze， E.和M. Kirschner, 1986。[j] .中国生物医学工程学报，2016,32(2):391 - 391。与动态不稳定模型一致（Mitchison， T.和M. Kirschner. 1984）。自然(Lond)。（312:237-242），在间期，MTs发生快速（2.6微米/分钟）的远端生长，同时出现新的中心体成核。大部分细胞质MT复合体在15-30分钟内被替换。在有丝分裂期间，星体MT显示相同的更新模式，但MT系统的更新要快得多（大约6分钟）。我们集中研究了前期着丝点纤维的结构，并观察到：(a)在前期和前期中期早期，只要着丝点还没有被mt连接到极点，就不会看到微管蛋白在着丝点附近的结合；(b)近端时间依赖性结合只发生在从中心体发出的预先存在的着丝粒mt中。因此，在未受干扰的前中期细胞中，着丝点可能不作为独立的成核位点。这证实了一个模型，在前期中期，快速探测的中心体mt被着丝点抓住，然后发生微管蛋白结合。
Microtubule (MT) dynamics in PtK2 cells have been investigated using in vivo injection of unmodified Paramecium ciliary tubulin and time-lapse fixation. The sites of incorporation of the axonemal tubulin were localized using a specific antibody which does not react with vertebrate cytoplasmic tubulin (Adoutte, A., M. Claisse, R. Maunoury, and J. Beisson. 1985. J. Mol. Evol. 22:220-229), followed by immunogold labeling, Nanovid microscopy, and ultrastructural observation of the same cells. We confirm data from microinjection of labeled tubulins in other cell types (Soltys, B. J., and G. G. Borisy. 1985. J. Cell Biol. 100:1682-1689; Mitchison, T., L. Evans, E. Schulze, and M. Kirschner. 1986. Cell. 45:515-527; Schulze, E., and M. Kirschner. 1986. J. Cell Biol. 102:1020-1031). In agreement with the dynamic instability model (Mitchison, T., and M. Kirschner. 1984. Nature (Lond.). 312:237-242), during interphase, fast (2.6 microns/min) distal growth of MTs occurs, together with new centrosomal nucleation. Most of the cytoplasmic MT complex is replaced within 15-30 min. During mitosis, astral MTs display the same pattern of renewal, but the turnover of the MT system is much faster (approximately 6 min). We have concentrated on the construction of the kinetochore fibers during prometaphase and observe that (a) incorporation of tubulin in the vicinity of the kinetochores is not seen during prophase and early prometaphase as long as the kinetochores are not yet connected to a pole by MTs; (b) proximal time- dependent incorporation occurs only into preexisting kinetochore MTs emanating from centrosomes. Consequently, in undisturbed prometaphase cells, the kinetochores probably do not act as independent nucleation sites. This confirms a model in which, at prometaphase, fast probing centrosomal MTs are grabbed by the kinetochores, where tubulin incorporation then takes place.
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DOI：
10.1083/jcb.84.3.717
发表时间：
1980-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Adoutte A;Ramanathan R;Lewis RM;Dute RR;Ling KY;Kung C;Nelson DL
通讯作者：
Nelson DL
DOI：
10.1083/jcb.102.3.1007
发表时间：
1986-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kristofferson D;Mitchison T;Kirschner M
通讯作者：
Kirschner M
影响因子：
3.9
作者：
ADOUTTE, A;CLAISSE, M;BEISSON, J
通讯作者：
BEISSON, J
影响因子：
5.6
作者：
FARRELL, KW;WILSON, L
通讯作者：
WILSON, L
影响因子：
2.7
作者：
KURIYAMA, R
通讯作者：
KURIYAMA, R