Yamauchi Seiji, Yamanaka Kunitoshi, Ogura Teru
Division of Molecular Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
Biochem Biophys Res Commun. 2006 Jun 30;345(2):746-53. doi: 10.1016/j.bbrc.2006.04.160. Epub 2006 May 5.
Caenorhabditis elegans possesses two p97/VCP/Cdc48p homologues, named CDC-48.1 (C06A1.1) and CDC-48.2 (C41C4.8), although their expression regulation and functional diversity have not yet been studied. We therefore investigated spatial and temporal expression patterns of two p97 homologues in this study. RT-PCR and Western blot analysis showed that the amount of cdc-48.1 was about twofold of that of cdc-48.2 in adults and that two p97 homologues were induced by ER stress. The amount of cdc-48.1 mRNA did not increase in the cdc-48.2 deletion mutant and vice versa. In situ hybridization showed that two p97 homologues are mainly expressed in germ cells. In vivo expression analysis by using GFP translational fusion constructs revealed that CDC-48.1::GFP was expressed from embryos through to adult worms, while CDC-48.2::GFP was expressed mainly in embryos. These results suggest that the expression of two p97 homologues of C. elegans is differently regulated and independent of each other.
秀丽隐杆线虫拥有两个p97/VCP/Cdc48p同源物,分别命名为CDC-48.1(C06A1.1)和CDC-48.2(C41C4.8),不过它们的表达调控和功能多样性尚未得到研究。因此,我们在本研究中调查了这两个p97同源物的时空表达模式。逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析表明,在成虫中cdc-48.1的量约为cdc-48.2的两倍,并且这两个p97同源物受到内质网应激的诱导。在cdc-48.2缺失突变体中cdc-48.1 mRNA的量没有增加,反之亦然。原位杂交显示这两个p97同源物主要在生殖细胞中表达。通过使用绿色荧光蛋白(GFP)翻译融合构建体进行的体内表达分析表明,CDC-48.1::GFP从胚胎期到成虫期均有表达,而CDC-48.2::GFP主要在胚胎期表达。这些结果表明,秀丽隐杆线虫的两个p97同源物的表达受到不同的调控且相互独立。