Cdc42Hs facilitates cytoskeletal reorganization and neurite outgrowth by localizing the 58-kD insulin receptor substrate to filamentous actin.
Cdc42Hs facilitates cytoskeletal reorganization and neurite outgrowth by localizing the 58-kD insulin receptor substrate to filamentous actin.
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DOI：
10.1083/jcb.152.3.579
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发表时间：
2001-02-05
期刊：
影响因子：
--
通讯作者：
Ahmed S
中科院分区：
文献类型：
--
作者：
Govind S;Kozma R;Monfries C;Lim L;Ahmed S
Cdc42Hs参与细胞骨架重组，是N1E-115细胞中神经突生长所必需的。为了研究Cdc42Hs调节这些过程的分子机制，通过酵母双杂交试验寻找新的Cdc42Hs蛋白伴侣。在这里，我们确定的58 kD底物的胰岛素受体酪氨酸激酶（IRS-58）作为Cdc42Hs的目标。IRS-58是脑富集蛋白，其包含至少四个蛋白质-蛋白质相互作用位点：Cdc42Hs结合位点、Src同源性（SH）3-结合位点、SH 3结构域和色氨酸、色氨酸（WW）-结合结构域。IRS-58在Swiss 3T3细胞中的表达导致丝状（F）-肌动蛋白细胞骨架的重组，涉及应力纤维的损失以及丝状伪足和簇的形成。在N1E-115细胞中，IRS-58诱导高度复杂的神经突生长。IRS-58缺失突变体的表达，它缺乏SH3-和WW-binding域，诱导N1E-115细胞中的神经突延伸而不复杂。在Swiss 3 T3细胞和N1 E-115细胞中，IRS-58与F-肌动蛋白共定位在簇和丝状伪足中。IRS-581267 N突变体不能结合Cdc42Hs未能定位与F-肌动蛋白诱导神经突生长或显着的细胞骨架重组。这些结果表明，Cdc42Hs促进细胞骨架重组和神经突生长的本地化蛋白复合物通过衔接蛋白，如IRS-58的F-肌动蛋白。
Cdc42Hs is involved in cytoskeletal reorganization and is required for neurite outgrowth in N1E-115 cells. To investigate the molecular mechanism by which Cdc42Hs regulates these processes, a search for novel Cdc42Hs protein partners was undertaken by yeast two-hybrid assay. Here, we identify the 58-kD substrate of the insulin receptor tyrosine kinase (IRS-58) as a Cdc42Hs target. IRS-58 is a brain-enriched protein comprising at least four protein–protein interaction sites: a Cdc42Hs binding site, an Src homology (SH)3-binding site, an SH3 domain, and a tryptophan, tyrptophan (WW)-binding domain. Expression of IRS-58 in Swiss 3T3 cells leads to reorganization of the filamentous (F)-actin cytoskeleton, involving loss of stress fibers and formation of filopodia and clusters. In N1E-115 cells IRS-58 induces neurite outgrowth with high complexity. Expression of a deletion mutant of IRS-58, which lacks the SH3- and WW-binding domains, induced neurite extension without complexity in N1E-115 cells. In Swiss 3T3 cells and N1E-115 cells, IRS-58 colocalizes with F-actin in clusters and filopodia. An IRS-581267N mutant unable to bind Cdc42Hs failed to localize with F-actin to induce neurite outgrowth or significant cytoskeletal reorganization. These results suggest that Cdc42Hs facilitates cytoskeletal reorganization and neurite outgrowth by localizing protein complexes via adaptor proteins such as IRS-58 to F-actin.
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影响因子：
5.3
作者：
Manser, E;Huang, HY;Lim, L
通讯作者：
Lim, L
影响因子：
5.3
作者：
Abbott, MA;Wells, DG;Fallon, JR
通讯作者：
Fallon, JR
影响因子：
5.3
作者：
Kozma, R;Sarner, S;Lim, L
通讯作者：
Lim, L
影响因子：
64.8
作者：
MANSER, E;LEUNG, T;LIM, L
通讯作者：
LIM, L
影响因子：
64.8
作者：
MANSER, E;LEUNG, T;LIM, L
通讯作者：
LIM, L