Jan E, Motzny C K, Graves L E, Goodwin E B
Department of Cell and Molecular Biology and Lurie Cancer Center, Northwestern University Medical School, Chicago, IL 60611, USA.
EMBO J. 1999 Jan 4;18(1):258-69. doi: 10.1093/emboj/18.1.258.
The Caenorhabditis elegans sex determination gene, tra-2, is translationally regulated by elements in the 3'-untranslated region called TGEs. TGEs govern the translation of mRNAs in both invertebrates and vertebrates, indicating that this is a highly conserved mechanism for controlling gene activity. A factor called DRF, found in worm extracts binds the TGEs and may be a repressor of translation. Using the yeast three-hybrid screen and RNA gel shift analysis, we have found that the protein GLD-1, a germline-specific protein and a member of the STAR family of RNA-binding proteins, specifically binds to the TGEs. GLD-1 is essential for oogenesis, and is also necessary for spermatogenesis and inhibition of germ cell proliferation. Several lines of evidence demonstrate that GLD-1 is a translational repressor acting through the TGEs to repress tra-2 translation. GLD-1 can repress the translation of reporter RNAs via the TGEs both in vitro and in vivo, and is required to maintain low TRA-2A protein levels in the germline. Genetic analysis indicates that GLD-1 acts upstream of the TGE control. Finally, we show that endogenous GLD-1 is a component of DRF. The conservation of the TGE control and the STAR family suggests that at least a subset of STAR proteins may work through the TGEs to control translation.
秀丽隐杆线虫的性别决定基因tra-2在3'非翻译区中被称为TGEs的元件进行翻译调控。TGEs调控无脊椎动物和脊椎动物中mRNA的翻译,这表明这是一种高度保守的控制基因活性的机制。在蠕虫提取物中发现的一种名为DRF的因子能结合TGEs,可能是翻译的抑制因子。通过酵母三杂交筛选和RNA凝胶迁移分析,我们发现蛋白质GLD-1,一种生殖系特异性蛋白且是RNA结合蛋白STAR家族的成员,能特异性地结合TGEs。GLD-1对卵子发生至关重要,对精子发生和抑制生殖细胞增殖也必不可少。多条证据表明GLD-1是一种通过TGEs起作用来抑制tra-2翻译的翻译抑制因子。GLD-1在体外和体内都能通过TGEs抑制报告RNA的翻译,并且在生殖系中维持低水平的TRA-2A蛋白是必需的。遗传分析表明GLD-1在TGE控制的上游起作用。最后,我们表明内源性GLD-1是DRF的一个组成部分。TGE控制和STAR家族的保守性表明,至少一部分STAR蛋白可能通过TGEs来控制翻译。