FUNCTIONS OF A CONSERVED GERMLINE HELICASE IN C.ELEGANS

描述（由申请人提供）：真核生物似乎各自编码一个单一的 CGH- 1（保守的生殖系解旋酶）的直系同源物，CGH-1是一种DEAD盒RNA解旋酶， 在许多生物体中是种系特异性的，并且是酵母中配子发生所必需的。 我们证明了C.秀丽线虫CGH-1是生殖细胞特异性的， 胚胎，并与生殖系P颗粒和其他假定的 mRNA-蛋白质颗粒。cgh-1是形成功能性配子所必需的， 如果没有它，基本上所有的卵母细胞都会通过生殖细胞特异性的细胞凋亡而死亡。 通路这种机制通常会杀死一半的发育中的卵母细胞， 显然是为了将它们的细胞质提供给它们幸存的姐妹篇。我们 研究结果表明，生理生殖细胞凋亡，一个保守的方面， 配子发生，可以作为一种监测机制，监测各方面的 生殖细胞质形成我们假设CGH- 1调节代谢 或在配子发生期间储存的特定mRNA的翻译，以及 这些功能的阐明将产生新的和广泛适用的 生殖细胞发育和细胞凋亡的见解。我们将测试这些模型， 阐明CGH-I如何有助于配子发生的关键方面，和 在分子水平上发挥作用。在后面的实验中，我们将阐明 其复杂表达模式的基础，决定它是否与 在功能上与RNA相关的复合物结合， 物种，测试它是否与母体mRNA相关，并使用无偏 鉴定其他可能具有物种特异性的CGH-1结合蛋白的方法。 这些实验将提供关于细胞凋亡是如何被触发的见解， 发展中的C.线虫卵母细胞，可能广泛适用于了解 生殖细胞生理死亡的原因与此同时， 阐明转录后基因调控机制，可能是 在配子发生过程中的保守重要性，并可能为增殖或 其他类型干细胞的存活。

DESCRIPTION (provided by applicant): Eukaryotes each appear to encode a single ortholog of CGH- 1 (conserved germline helicase), a DEAD box RNA helicase that is germline-specific in many organisms and required for gametogenesis in yeast. We have shown that C. elegans CGH-1 is specific to the germline and early embryo, and is associated with germline P granules and other putative mRNA-protein particles. cgh-1 is required to form functional gametes, and without it essentially all oocytes die by apoptosis through a germline-specific pathway. This mechanism normally kills up to half of all developing oocytes, apparently so that their cytoplasm is provided to their surviving sisters. Our findings suggest that physiological germline apoptosis, a conserved aspect of gametogenesis, can act as a surveillance mechanism that monitors aspects of germline cytoplasm formation. We hypothesize that CGH- 1 regulates metabolism or translation of particular mRNAs that are stored during gametogenesis, and that elucidation of these functions will yield novel and broadly applicable insights into germline development and apoptosis. We will test these models by elucidating critical aspects of how CGH-l contributes to gametogenesis, and functions at the molecular level. In the latter experiments, we will elucidate the basis for its complex expression pattern, determine whether it interacts functionally with RNA-associated complexes that bind its orthologs in other species, test whether it associates with maternal mRNAs, and use unbiased methods to identify other CGH-1-binding proteins that may be species-specific. These experiments will provide insights into how apoptosis is triggered in developing C. elegans oocytes that may be broadly applicable to understanding why physiological germline cell death occurs. At the same time, they will elucidate post-transcriptional gene regulation mechanisms that may be of conserved importance during gametogenesis, and possibly for proliferation or survival of other stem cell types.