Andrä Kerstin, Kornacker Iris, Jörgl Almut, Zörer Michael, Spazierer Daniel, Fuchs Peter, Fischer Irmgard, Wiche Gerhard
Institute of Biochemistry and Molecular Cell Biology, Vienna Biocenter, University of Vienna, Austria.
J Invest Dermatol. 2003 Feb;120(2):189-97. doi: 10.1046/j.1523-1747.2003.12027.x.
The various plectin isoforms are among the major crosslinking elements of the cytoskeleton. The importance of plectin in epithelia is convincingly supported by the severe skin blistering observed in plectin-deficient humans and mice. Here, we identified plectin 1a (> 500 kDa), a full length plectin variant containing the sequence encoded by the alternative first exon 1a, as the isoform most prominently expressed in human and mouse keratinocytes. In skin sections and cultured keratinocytes, plectin 1a was shown to colocalize with hemidesmosomal structures. In contrast, a second isoform expressed in epithelia, plectin 1c, differing from 1a merely by a short N-terminal sequence, colocalized with microtubules. Expression of plectin 1a, but not of its N-terminal fragment alone, or of a third alternative full length isoform (plectin 1), restored the reduced number of hemidesmosome-like stable anchoring contacts in cultured plectin-null keratinocytes. Our results show for the first time that different isoforms of a cytolinker protein expressed in one cell type perform distinct functions. Moreover, the identification of plectin 1a as the isoform defects in which cause skin blistering in plectin-related genetic diseases, such as epidermolysis bullosa simplex MD and epidermolysis bullosa simplex Ogna, could have implications for the future development of clinical therapies for patients.
多种网蛋白异构体是细胞骨架的主要交联元件。网蛋白在表皮中的重要性在网蛋白缺陷的人类和小鼠中观察到的严重皮肤水疱现象中得到了令人信服的支持。在此,我们鉴定出网蛋白1a(>500 kDa),这是一种全长网蛋白变体,包含由选择性的第一个外显子1a编码的序列,是在人类和小鼠角质形成细胞中最显著表达的异构体。在皮肤切片和培养的角质形成细胞中,网蛋白1a显示与半桥粒结构共定位。相比之下,在上皮中表达的第二种异构体网蛋白1c,与1a仅在短的N端序列上不同,与微管共定位。网蛋白1a的表达,而不是单独的其N端片段或第三种选择性全长异构体(网蛋白1)的表达,恢复了培养的无网蛋白角质形成细胞中半桥粒样稳定锚定连接数量的减少。我们的结果首次表明,在一种细胞类型中表达的细胞连接蛋白的不同异构体执行不同的功能。此外,将网蛋白1a鉴定为在与网蛋白相关的遗传性疾病如单纯性大疱性表皮松解症MD和单纯性大疱性表皮松解症Ogna中导致皮肤水疱的异构体缺陷,可能对未来患者临床治疗的发展具有启示意义。