Keratin-containing inclusions affect cell morphology and distribution of cytosolic cellular components.
Keratin-containing inclusions affect cell morphology and distribution of cytosolic cellular components.
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含角蛋白的内含物影响细胞形态和细胞质细胞成分的分布。
DOI：
10.1016/j.yexcr.2004.12.009
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Sata,Michio
中科院分区：
文献类型：
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作者：
Hanada,Shinichiro;Harada,Masaru;Kumemura,Hiroto;Omary,MBishr;Kawaguchi,Takumi;Taniguchi,Eitaro;Koga,Hironori;Yoshida,Takafumi;Maeyama,Michiko;Baba,Shinji;Ueno,Takato;Sata,Michio
许多神经退行性疾病的特征在于存在与中间丝（IF）捆绑的蛋白质聚集体，并且在各种肝脏疾病中观察到称为马洛里体（MB）的类似结构。IF锚定在桥粒和半桥粒，然而，与其他细胞间连接的相互作用尚未确定。我们研究了IF包涵体对各种培养细胞中连接相关蛋白和胞浆蛋白的影响。我们在培养的细胞中进行了绿色荧光蛋白（GFP）标记的细胞角蛋白（CK）18突变体arg 89 cys（GFP-CK 18 R89 C）的基因转染，并观察到CK聚集以及IF网络的丢失。在各种连接相关蛋白中，在免疫荧光分析中，闭合小带-1和β-连环蛋白与CK聚集体共定位。对胞质蛋白、14-3-3-β-D蛋白、葡萄糖-6-磷酸脱氢酶和DsRed的免疫染色获得了类似的结果。E-cadherin是极化上皮细胞中的基底外侧膜蛋白，在GFP-CK 18 R89 C转染的细胞中存在于顶端和基底外侧域。此外，含有CK聚集体的细胞显著大于GFP标记的野生型CK 18（GFP-WT CK 18）转染或未转染的细胞（P < 0.01），并且有时它们的形态显著改变。我们的数据表明，CK聚集体不仅影响细胞形态，而且还影响各种胞质组分的定位，这可能会影响细胞功能。
Many neurodegenerative diseases are characterized by the presence of protein aggregates bundled with intermediate filaments (IFs) and similar structures, known as Mallory bodies (MBs), are observed in various liver diseases. IFs are anchored at desmosomes and hemidesmosomes, however, interactions with other intercellular junctions have not been determined. We investigated the effect of IF inclusions on junction-associated and cytosolic proteins in various cultured cells. We performed gene transfection of the green fluorescent protein (GFP)-tagged cytokeratin (CK) 18 mutant arg89cys (GFP-CK18 R89C) in cultured cells and observed CK aggregations as well as loss of IF networks. Among various junction-associated proteins, zonula occludens-1 and β-catenin were colocalized with CK aggregates on immunofluorescent analyses. Similar results were obtained on immunostaining for cytosolic proteins, 14-3-3 ζ protein, glucose-6-phosphate dehydrogenase and DsRed. E-cadherin, a basolateral membrane protein in polarized epithelia, was present on both the apical and basolateral domains in GFP-CK18 R89C-transfected cells. Furthermore, cells containing CK aggregates were significantly larger than GFP-tagged wild type CK18 (GFP-WT CK18)-transfected or non-transfected cells (P < 0.01) and sometimes their morphology was significantly altered. Our data indicate that CK aggregates affect not only cell morphology but also the localization of various cytosolic components, which may affect the cellular function.