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秀丽隐杆线虫WASP相互作用蛋白同源物WIP-1通过维持WSP-1蛋白水平参与形态发生。

Caenorhabditis elegans WASP-interacting protein homologue WIP-1 is involved in morphogenesis through maintenance of WSP-1 protein levels.

作者信息

Sawa Mariko, Takenawa Tadaomi

机构信息

Department of Biochemistry, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.

出版信息

Biochem Biophys Res Commun. 2006 Feb 10;340(2):709-17. doi: 10.1016/j.bbrc.2005.12.056. Epub 2005 Dec 20.

Abstract

Mammalian WASP and N-WASP are involved in reorganization of the actin cytoskeleton through activation of the Arp2/3 complex and in regulation of cell motility or cell shape changes. In the present study, we identified WASP-interacting protein homologue (WIP)-1 in Caenorhabditis elegans. WIP-1 contains the domains and sequences conserved among mammalian WIP family proteins. Yeast two-hybrid analysis detected a physical interaction between WIP-1 and WSP-1, the sole homologue of WASP/N-WASP in C. elegans. Western analysis of embryo lysates showed that RNA interference (RNAi) treatment for wip-1 decreased levels of WSP-1 protein, and wsp-1(RNAi) treatment decreased levels of WIP-1 protein. However, wsp-1 mRNA levels were not decreased in wip-1(RNAi)-treated embryos, and wip-1 mRNA levels were not decreased in wsp-1(RNAi)-treated embryos. Furthermore, disruption of WIP-1 by RNAi resulted in embryonic lethality with morphologic defects in hypodermal cell migration, a process known as ventral enclosure. This phenotype was similar to that observed in RNAi experiments for wsp-1. Immunostaining showed that WIP-1 was expressed by migrating hypodermal cells, as was WSP-1. This expression during ventral enclosure was reduced in wip-1(RNAi)-treated embryos and wsp-1(RNAi)-treated embryos. Our results suggest that C. elegans WIP-1 may function in hypodermal cell migration during ventral enclosure by maintaining levels of WSP-1.

摘要

哺乳动物的WASP和N-WASP通过激活Arp2/3复合物参与肌动蛋白细胞骨架的重组,并参与细胞运动或细胞形状变化的调节。在本研究中,我们在秀丽隐杆线虫中鉴定出WASP相互作用蛋白同源物(WIP)-1。WIP-1包含在哺乳动物WIP家族蛋白中保守的结构域和序列。酵母双杂交分析检测到WIP-1与WSP-1之间存在物理相互作用,WSP-1是秀丽隐杆线虫中WASP/N-WASP的唯一同源物。对胚胎裂解物的蛋白质免疫印迹分析表明,对wip-1进行RNA干扰(RNAi)处理会降低WSP-1蛋白水平,而对wsp-1进行RNAi处理会降低WIP-1蛋白水平。然而,在经wip-1(RNAi)处理的胚胎中,wsp-1 mRNA水平并未降低,在经wsp-1(RNAi)处理的胚胎中,wip-1 mRNA水平也未降低。此外,通过RNAi破坏WIP-1会导致胚胎致死,并伴有皮下细胞迁移的形态学缺陷,这一过程称为腹侧包被。该表型与在wsp-1的RNAi实验中观察到的相似。免疫染色显示,WIP-1由迁移的皮下细胞表达,WSP-1也是如此。在腹侧包被过程中的这种表达在经wip-1(RNAi)处理的胚胎和经wsp-1(RNAi)处理的胚胎中均降低。我们的结果表明,秀丽隐杆线虫的WIP-1可能通过维持WSP-1的水平在腹侧包被期间的皮下细胞迁移中发挥作用。

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