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14-3-3家族成员在非洲爪蟾发育中的差异作用。

Differential role of 14-3-3 family members in Xenopus development.

作者信息

Lau Jeffrey M C, Wu Chunlai, Muslin Anthony J

机构信息

Center for Cardiovascular Research, Department of Medicine, and Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

出版信息

Dev Dyn. 2006 Jul;235(7):1761-76. doi: 10.1002/dvdy.20816.

Abstract

The 14-3-3 proteins are intracellular dimeric phosphoserine/threonine binding molecules that participate in signal transduction, checkpoint control, nutrient sensing, and cell survival pathways. Previous work established that 14-3-3 proteins are required in early Xenopus laevis development by modulating fibroblast growth factor signaling. Although this general requirement for 14-3-3 proteins in Xenopus early embryogenesis is established, there is no information about the specific role of individual 14-3-3 genes. Botanical studies previously demonstrated functional specificity among 14-3-3 genes during plant development. In this study, an antisense morpholino oligo microinjection approach was used to characterize the requirement for six specific 14-3-3 family members in Xenopus embryogenesis. Microinjection experiments followed by Western blot analysis showed that morpholinos reduced specific 14-3-3 protein levels. Embryos lacking specific 14-3-3 isoforms displayed unique phenotypic defects. In particular, reduction of 14-3-3 tau (tau) protein, and to a lesser extent, 14-3-3 epsilon (epsilon), resulted in embryos with prominent gastrulation and axial patterning defects and reduced mesodermal marker gene expression. In contrast, reduction of 14-3-3 zeta (zeta) protein caused no obvious phenotypic abnormalities. Reduction of 14-3-3 gamma (gamma) protein resulted in eye defects without gastrulation abnormalities. Therefore, individual 14-3-3 genes have separable functions in vertebrate embryonic development.

摘要

14-3-3蛋白是细胞内的二聚体磷酸丝氨酸/苏氨酸结合分子,参与信号转导、关卡控制、营养感知和细胞存活途径。先前的研究表明,14-3-3蛋白通过调节成纤维细胞生长因子信号通路,在非洲爪蟾早期发育中发挥作用。虽然14-3-3蛋白在非洲爪蟾早期胚胎发育中的总体需求已得到证实,但关于单个14-3-3基因的具体作用尚无相关信息。植物学研究先前已证明14-3-3基因在植物发育过程中具有功能特异性。在本研究中,采用反义吗啉代寡核苷酸显微注射方法,来确定非洲爪蟾胚胎发育中6个特定14-3-3家族成员的需求。显微注射实验及随后的蛋白质免疫印迹分析表明,吗啉代寡核苷酸降低了特定14-3-3蛋白的水平。缺乏特定14-3-3亚型的胚胎表现出独特的表型缺陷。特别是,14-3-3 tau(tau)蛋白水平降低,以及程度较轻的14-3-3 epsilon(epsilon)蛋白水平降低,导致胚胎出现明显的原肠胚形成和轴向模式缺陷,以及中胚层标记基因表达减少。相比之下,14-3-3 zeta(zeta)蛋白水平降低未引起明显的表型异常。14-3-3 gamma(gamma)蛋白水平降低导致眼部缺陷,但没有原肠胚形成异常。因此,单个14-3-3基因在脊椎动物胚胎发育中具有可分离的功能。

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