Goossens Steven, Janssens Barbara, Vanpoucke Griet, De Rycke Riet, van Hengel Jolanda, van Roy Frans
Department for Molecular Biomedical Research, VIB-Ghent University, Technologiepark 927, B-9052 Ghent, Belgium.
FASEB J. 2007 Mar;21(3):647-55. doi: 10.1096/fj.06-6066com. Epub 2006 Dec 21.
AlphaT-catenin is a recently identified member of the alpha-catenin family of cell-cell adhesion molecules. For decades it was thought that alpha-catenins mediate solid cell-cell adhesion by linking the cadherin-mediated cell-cell adhesion complex with the actin cytoskeleton. However, the roles of alpha-catenins in this classical adhesion model have been questioned recently. AlphaT-catenin has a restricted expression pattern, in contrast to the ubiquitously expressed alphaE-catenin. High levels of alphaT-catenin were detected in heart and testis. Northern and Western blot experiments indicated that besides the standard full-length alphaT-catenin transcript, smaller alternative transcripts are expressed in testis. We report the cloning of two alternative transcripts of the mouse alphaT-catenin gene (transcript-B and -X), both of which are expressed in a testis-restricted manner from two putative alternative promoters. Alternative transcript-X encodes a smaller protein, isoform-X, which lacks the amino-terminal beta-catenin binding domain of the standard mouse alphaT-catenin protein, and is therefore unable to restore cell-cell adhesion in an alpha-catenin-negative colon carcinoma cell line. Immunohistochemical analysis showed specific localization of the alphaT-catenin isoform-X in the differentiating germ cells. In contrast to the standard full-length alphaT-catenin protein, this shortened isoform-X can bind to l-afadin, an important component of the nectin/afadin/ponsin adhesion complex that reportedly is essential for spermatogenesis.
αT-连环蛋白是细胞间黏附分子α-连环蛋白家族中最近发现的成员。几十年来,人们一直认为α-连环蛋白通过将钙黏蛋白介导的细胞间黏附复合物与肌动蛋白细胞骨架相连来介导实体细胞间黏附。然而,α-连环蛋白在这种经典黏附模型中的作用最近受到了质疑。与广泛表达的αE-连环蛋白不同,αT-连环蛋白具有受限的表达模式。在心脏和睾丸中检测到高水平的αT-连环蛋白。Northern印迹和Western印迹实验表明,除了标准的全长αT-连环蛋白转录本外,睾丸中还表达较小的可变转录本。我们报道了小鼠αT-连环蛋白基因的两种可变转录本(转录本-B和-X)的克隆,这两种转录本均从两个假定的可变启动子以睾丸特异性方式表达。可变转录本-X编码一种较小的蛋白质,即异构体-X,它缺乏标准小鼠αT-连环蛋白的氨基末端β-连环蛋白结合结构域,因此无法在α-连环蛋白阴性的结肠癌细胞系中恢复细胞间黏附。免疫组织化学分析显示αT-连环蛋白异构体-X在分化的生殖细胞中特异性定位。与标准的全长αT-连环蛋白不同,这种缩短的异构体-X可以与l-afadin结合,l-afadin是nectin/afadin/ponsin黏附复合物的重要组成部分,据报道对精子发生至关重要。