Elucidation of the protein phosphorylation signaling and the regulatory mechanism of cytoskeleton

阐明蛋白质磷酸化信号和细胞骨架的调控机制

基本信息

批准号：
18370085
负责人：
INAGAKI Masaki
金额：
$ 11.28万
依托单位：
Aichi Cancer Center Research Institute
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18370085/
关键词：
protein phosphorylation cell cycle cytoskeleton cell adhesion cell polarity intermediate filament centrosome ケラチン

项目摘要

We recently reported Chk1 to be phosphorylated at Ser286 and Ser301 by cyclin-dependent kinase(Cdk) 1 during mitosis. In order to elucidate functional consequences, we produced a rat monoclonal antibody which specifically recoginzes Chk1 phosphorylated at Ser301. Immunocytochemical analyses using this antibody revealed Chk1-Ser301 phosphorylation from entry into mitosis(prophase). The level of phosphorylation appeared to peak at metaphase and to gradually decrease after anaphase. In prophase when chromosome condensation occurs without nuclear envelope breakdown, we observed two phosphorylation patterns. At earlier stages, the phophorylation tended to be observed mainly in the nucleus. However, at later stages of prophase, Chk1 phosphorylated at Ser301 appealed to exist mainly in the cytoplasm. This translocation was inhibited by treatment with leptomycin B, which blocks Crm1-mediated nuclear export. Induced expression of S286A/S301A mutant caused significant delay in mitotic entry, com … More pared with the WT case. In S286A/S301A mutant prophase cells, staining was observed mainly in the nucleus although in WT cells location was also found in the cytoplasm. These results suggested that Cdk1 controls the cytoplasmic sequestration of Chk1 through the phosphorylation of Chk1.We recently identified two novel keratin filament-binding proteins : trichoplein and Albatross. Trichoplein was found to localize not only on keratin filaments but also on mother centnoles. Knockdown in HeLa cells revealed that trichoplein is essential for anchoring of microtubules and ninein on appendages of mother centrioles. Albatross localizes not only on keratin filaments but also in the vicinity of the apical junctional complex(AJC), a cell-cell adhesive apparatus composed of tight junctions, adherens junctions and desmosomes. Knockdown in A549, lung adenocarcinoma, cells revealed that Albatross regulates the formation of AJC and lateral domains in cells. In addition, we found that keratin promotes this function, using keratin-rescued SW13 cells. Less

我们最近报道了CHK1在有丝分裂的情况下通过cyclin依赖性激酶（CDK）1在Ser286和Ser301上进行磷酸化。 SE在预言中，染色体凝结的情况下，在核酸杆菌中，phophorylation均倾向于在核的后期中观察到。 B，阻止CRM1介导的S286A/S301A的核心，com在S286A/S301A突变体中的WT情况更加严重。细胞质。这些结果表明，CDK1通过CHK1的细胞质隔离。我们发现蛋白质结合蛋白：毛状蛋白质是在Hela细胞中均未固定trichoplein trichodin not trichoplein。中心元素。使用角蛋白验证的SW13细胞。