LOCALIZATION OF NEWLY SYNTHESIZED VIMENTIN SUBUNITS REVEALS A NOVEL MECHANISM OF INTERMEDIATE FILAMENT ASSEMBLY
LOCALIZATION OF NEWLY SYNTHESIZED VIMENTIN SUBUNITS REVEALS A NOVEL MECHANISM OF INTERMEDIATE FILAMENT ASSEMBLY
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DOI：
10.1016/0092-8674(90)90593-4
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发表时间：
1990-02-09
期刊：
影响因子：
64.5
通讯作者：
LAZARIDES, E
中科院分区：
文献类型：
--
作者：
NGAI, J;COLEMAN, TR;LAZARIDES, E
我们已经评估了中间丝组装的机制，通过检测在转染的小鼠成纤维细胞中诱导型启动子控制下表达的鸡波形蛋白亚基的掺入位点。新合成的波形蛋白的本地化确定在短时间内诱导合成的免疫荧光和免疫电子显微镜，使用鸡波形蛋白特异性抗体。在既不影响可溶性亚基池也不影响内源性丝的稳态分布的条件下，新合成的波形蛋白在整个细胞的许多离散位点并入波形蛋白丝网络中。随着时间的推移，新组装的波形蛋白的图案转化为与预先存在的波形蛋白细丝一致的连续阵列。这些结果是一致的一种新的机制，中间丝组装，从而发生生长的中间丝的地形限制和本地化的亚基添加，需要一个短暂的破坏丝的完整性。
We have assessed the mechanism of intermediate filament assembly by assaying the sites of incorporation of chiken vimentin subunits expressed under the control of an inducible promoter in transfected mouse fibroblasts. The localization of newly synthetized vimentin was determined by immunofluorescence and immunoelectron microscopy at short time periods of induced synthesis, using antibodies specific for chicken vimentin. Under conditions where neither the soluble subunit pools nor the steady-state distribution of endogenous filaments are affected, newly synthesized vimentin incorporates into the vimentin filament network at numerous and discrete sites throughout the cell. Over time, the pattern of newly assembled vimentin converts to a continuous array coincident with preexisting vimentin filaments. These results are consistent with a novel mechanism of intermediate filament assembly, whereby growth of intermediate filaments occurs by topographically restricted and localized subunit addition, necessitating a transient disruption of filament integrity.