Cytoskeletal architecture of isolated mitotic spindle with special reference to microtubule-associated proteins and cytoplasmic dynein.
Cytoskeletal architecture of isolated mitotic spindle with special reference to microtubule-associated proteins and cytoplasmic dynein.
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分离的有丝分裂纺锤体的细胞骨架结构，特别参考微管相关蛋白和细胞质动力蛋白。
DOI：
10.1083/jcb.101.5.1858
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发表时间：
1985-11
影响因子：
7.8
通讯作者：
HISANAGA, S
中科院分区：
文献类型：
--
作者：
HIROKAWA, N;TAKEMURA, R;HISANAGA, S
我们研究了细胞骨架结构的分离有丝分裂器从海胆卵使用速冻，深蚀刻电子显微镜。这种方法揭示了纺锤体微管之间存在广泛的直的和分支的三维网络。纺锤体微管的表面几乎完全被六边形堆积的、小的、圆形纽扣状结构覆盖，这些结构的形状和大小非常均匀（直径约8 nm），并且这些微管纽扣经常为相邻微管之间的横桥提供基底。通过高盐（0.6 M NaCl）提取从微管表面去除这些结构。从有丝分裂纺锤体中分离出微管相关蛋白（MAPs）和微管（主要由大量的75 kD蛋白和一些高分子量（250 kD、245 kD）蛋白组成）。微管的表面完全覆盖着同样的六角形包装的圆形按钮，其排列是密切相关的微管蛋白二聚体。此外，还发现了一些短的横桥和一些长的横桥。高盐处理（0.6 M NaCl）提取75-kD蛋白质和高分子量蛋白质，并从微管表面去除微管按钮和大部分的横桥。考虑到从有丝分裂纺锤体中分离出的MAPs中75- kD蛋白含量相对较高，可以得出结论，这些微管按钮可能由75-kD MAP组成，并且体内的一些横桥可能属于MAPs。另一种颗粒，尺寸较大（直径11-26 nm），有时也与有丝分裂纺锤体的微管表面相关。一个很好的侧臂有时连接较大的颗粒与相邻的微管。用免疫细胞化学方法研究了海胆精子鞭毛21 S动力蛋白单克隆抗体（D57）和胶体金标记的第二抗体对细胞质动力蛋白ATP酶在有丝分裂纺锤体中的定位。免疫金颗粒与纺锤体微管密切相关。其中76%位于微管表面50 nm以内，55%位于微管表面20 nm以内。这些金颗粒在中期和后期的半纺锤体的极微管和动粒微管上都有零星分布。在后期，它们也定位于姐妹染色单体之间的微管上。这些数据表明，细胞质动力蛋白附着在海胆有丝分裂纺锤体的微管。（400字处截断摘要）
We have studied cytoskeletal architectures of isolated mitotic apparatus from sea urchin eggs using quick-freeze, deep-etch electron microscopy. This method revealed the existence of an extensive three- dimensional network of straight and branching crossbridges between spindle microtubules. The surface of the spindle microtubules was almost entirely covered with hexagonally packed, small, round button- like structures which were very uniform in shape and size (approximately 8 nm in diameter), and these microtubule buttons frequently provided bases for crossbridges between adjacent microtubules. These structures were removed from the surface of microtubules by high salt (0.6 M NaCl) extraction. Microtubule- associated proteins (MAPs) and microtubules isolated from mitotic spindles which were mainly composed of a large amount of 75-kD protein and some high molecular mass (250 kD, 245 kD) proteins were polymerized in vitro and examined by quick-freeze, deep-etch electron microscopy. The surfaces of microtubules were entirely covered with the same hexagonally packed round buttons, the arrangement of which is intimately related to that of tubulin dimers. Short crossbridges and some longer crossbridges were also observed. High salt treatment (0.6 M NaCl) extracted both 75-kD protein and high molecular weight proteins and removed microtubule buttons and most of crossbridges from the surface of microtubules. Considering the relatively high amount of 75- kD protein among MAPs isolated from mitotic spindles, it is concluded that these microtubule buttons probably consist of 75-kD MAP and that some of the crossbridges in vivo could belong to MAPs. Another kind of granule, larger in size (11-26 nm in diameter), was also on occasion associated with the surface of microtubules of mitotic spindles. A fine sidearm sometimes connected the larger granule to adjacent microtubules. Localization of cytoplasmic dynein ATPase in the mitotic spindle was investigated by electron microscopic immunocytochemistry with a monoclonal antibody (D57) against sea urchin sperm flagellar 21S dynein and colloidal gold-labeled second antibody. Immunogold particles were closely associated with spindle microtubules. 76% of these were within 50 nm and 55% were within 20 nm from the surface of the microtubules. These gold particles were sporadically found on both polar and kinetochore microtubules of half-spindles at both metaphase and anaphase. They localized also on the microtubules between sister chromatids in late anaphase. These data indicate that cytoplasmic dynein is attached to the microtubules in sea urchin mitotic spindles.(ABSTRACT TRUNCATED AT 400 WORDS)
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DOI：
10.1083/jcb.74.2.377
发表时间：
1977-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
McDonald K;Pickett-Heaps JD;McIntosh JR;Tippit DH
通讯作者：
Tippit DH
影响因子：
7.8
作者：
ERICKSON, HP
通讯作者：
ERICKSON, HP
影响因子：
2.7
作者：
HISANAGA, SI;SAKAI, H
通讯作者：
SAKAI, H
DOI：
10.1016/s0022-5320(83)90047-3
发表时间：
1983-01-01
期刊：
JOURNAL OF ULTRASTRUCTURE RESEARCH
影响因子：
--
作者：
MILLER, KR;PRESCOTT, CS;LASSIGNAL, NL
通讯作者：
LASSIGNAL, NL
DOI：
10.1083/jcb.93.3.788
发表时间：
1982-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Keller TC 3rd;Rebhun LI
通讯作者：
Rebhun LI