Plectin is a widely expressed protein that is very large in size and that has all the attributes of a multifunctional crosslinking and organizing element of the cytoskeleton. It displays a multidomain structure, versatile binding activities, and subcellular localizations that enable it to strengthen cells against mechanical stress forces. Moreover, hereditary gene defects in plectin cause epidermolysis bullosa simplex (EBS)-MD, a severe skin blistering disease with muscular dystrophy. Here we report the analysis of the exonintron organization of the rat plectin gene and the identification of several different isoforms on the transcriptional level. We show that of 35 coding exons identified, 4 serve as alternative first exons splicing into the same successive exon 2, which is the first of 7 exons encoding a highly conserved actin-binding domain. RNase protection mapping of transcripts containing 3 of the identified 4 alternate first exons revealed their coexpression in rat glioma C6 cells and in a series of different rat tissues that we examined. Significant variations in expression levels of first exons indicated the possibility of tissue-specific promoter usage. In addition, plectin splice variants lacking exon 31 (> 3 kb), which encodes the entire rod domain of the molecule, were identified in a variety of rat tissues. This study provides first insights into a complex plectin gene regulatory machinery with similarities to that of dystrophin.

译文

pletin是一种广泛表达的蛋白质,其大小非常大,并且具有细胞骨架的多功能交联和组织元件的所有属性。它显示出多域结构,多种结合活性和亚细胞定位,使其能够增强细胞抵抗机械应力的能力。此外,plectin的遗传性基因缺陷会导致大疱性表皮松解症 (EBS)-MD,这是一种伴有肌营养不良的严重皮肤水疱性疾病。在这里,我们报告了对大鼠plectin基因外显子组织的分析,并在转录水平上鉴定了几种不同的同工型。我们显示,在鉴定出的35个编码外显子中,有4个作为替代的第一个外显子拼接成相同的连续外显子2,这是编码高度保守的肌动蛋白结合域的7个外显子中的第一个。包含鉴定出的4个交替第一外显子中的3个的转录物的RNase保护作图显示了它们在大鼠神经胶质瘤C6细胞和我们检查的一系列不同大鼠组织中的共表达。第一个外显子表达水平的显着变化表明使用组织特异性启动子的可能性。此外,在多种大鼠组织中鉴定出缺少外显子31 (> 3 kb) 的pletin剪接变体,该外显子编码分子的整个杆结构域。这项研究提供了与肌营养不良蛋白相似的复杂pletin基因调控机制的第一个见解。

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