Effects of Mutant Keratin Genes in Transfected Epidermal Cells : Establishment of Cell Models for Genodermatoses

突变角蛋白基因对转染表皮细胞的影响：遗传性皮肤病细胞模型的建立

• 批准号: 07670966
• 负责人: MANABE Motomu
• 金额: $ 1.41万
• 依托单位: JUNTENDO UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670966/
• 关键词: keratins; keratin diseases; transfection

To define the interactions required for the filament assembly of differentiation-specific keratins, active copies of mouse hair keratin mHa1 and mHb4 genes were introduced into a rat kangaroo kidney epithelial cell line (PtK2) and a rat stratified squamous epithelial cell line (rat epidermal keratinocyte). In PtK2 transient transfectants when introduced individually or in combination, mHa1 and mHb4 formed aggregates of ring-like structures of various sizes at the perinuclear region with no evidence of organization into a keratin network. These aggregates altered the distribution of the endogenous keratins and vimentin. In most of the cells carrying the ring-like structures of mHa1 and mHb4 around the nucleus, the endogenous keratin network collapsed and localized around the nucleus. Furthermore, the densely accumulated endogenous keratin surrounded the ring-like aggregates with partial co-localization. However, when transfected into the rat epidermal keratinocytes, mHa1 and mHb4 were able to co-localize with the well-developed cytoskeleton of endogenous keratins. These results showed that, in contrast to keratin pairs K5/K14 and K8/K18, the mHa1/mHb4 pair is unable to develop an extensive keratin network on its own, and that there are possible differential abilities among these hair keratins and other keratins to form well-developed cytoplasmic networks.

为了确定分化特异性角蛋白丝组装所需的相互作用，将小鼠毛发角蛋白 mHa1 和 mHb4 基因的活性拷贝引入大鼠袋鼠肾上皮细胞系 (PtK2) 和大鼠分层鳞状上皮细胞系（大鼠表皮角质形成细胞）中。在 PtK2 瞬时转染子中，当单独或组合引入时，mHa1 和 mHb4 在核周区域形成各种大小的环状结构聚集体，没有组织成角蛋白网络的证据。这些聚集体改变了内源性角蛋白和波形蛋白的分布。在大多数细胞核周围携带 mHa1 和 mHb4 环状结构的细胞中，内源角蛋白网络崩溃并集中在细胞核周围。此外，密集积累的内源角蛋白以部分共定位的方式包围环状聚集体。然而，当转染到大鼠表皮角质形成细胞中时，mHa1 和 mHb4 能够与发育良好的内源性角蛋白细胞骨架共定位。这些结果表明，与角蛋白对 K5/K14 和 K8/K18 相比，mHa1/mHb4 对自身无法形成广泛的角蛋白网络，并且这些毛发角蛋白和其他角蛋白形成发育良好的细胞质网络的能力可能存在差异。

项目成果

Manabe M et al: "Assembly of hair keratins in transfected epithelial cells." Biochem.Biophys.Res.Comm.229. 965-973 (1996)

Manabe M 等人：“转染上皮细胞中毛发角蛋白的组装。”

Manabe M et al.: "Assembly of hair keratins in transfected epithelial cells." Biochem.Biophys.Res.Comm.229. 965-973 (1996)

Manabe M 等人：“转染上皮细胞中毛发角蛋白的组装。”