仓鼠结蛋白基因的特性：基因转移后非肌肉细胞中结蛋白丝的表达与形成

Characterization of the hamster desmin gene: expression and formation of desmin filaments in nonmuscle cells after gene transfer.

作者信息

Quax W, van den Broek L, Egberts W V, Ramaekers F, Bloemendal H

出版信息

Cell. 1985 Nov;43(1):327-38. doi: 10.1016/0092-8674(85)90038-8.

Abstract

The structural organization of the hamster gene encoding the intermediate filament (IF) protein desmin has been determined. The gene, 6.5 kb in length, contains nine exons with a total length of 2169 nucleotides. Remarkably, the intervening sequences map at positions that fully correspond to those of the vimentin gene. The derived complete primary structure for hamster desmin (468 amino acids; 53,250 daltons) reveals striking species variations in the NH2-terminal domain of desmin. A plasmid containing the complete transcription unit of the desmin gene was transfected into hamster lens cells and into human epithelial (HeLa) cells. In both nonmuscle cell lines the desmin gene was biologically active. The synthesized desmin assembled into authentic IFs, as monitored by immunofluorescence. Double immunofluorescence staining showed that the newly formed desmin filaments colocalize with preexisting vimentin filaments, but not with preexisting keratin filaments.

摘要

已确定了仓鼠中编码中间丝（IF）蛋白结蛋白的基因的结构组织。该基因长度为6.5kb，包含9个外显子，总长度为2169个核苷酸。值得注意的是，间隔序列定位在与波形蛋白基因的序列完全对应的位置。推导得到的仓鼠结蛋白的完整一级结构（468个氨基酸；53250道尔顿）显示，结蛋白的NH2末端结构域存在显著的物种差异。将含有结蛋白基因完整转录单位的质粒转染到仓鼠晶状体细胞和人上皮（HeLa）细胞中。在这两种非肌肉细胞系中，结蛋白基因均具有生物活性。通过免疫荧光监测，合成的结蛋白组装成了真正的中间丝。双重免疫荧光染色显示，新形成的结蛋白丝与预先存在的波形蛋白丝共定位，但不与预先存在的角蛋白丝共定位。