Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans.
Proximity labeling identifies LOTUS domain proteins that promote the formation of perinuclear germ granules in C. elegans.
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作者：
Price IF;Hertz HL;Pastore B;Wagner J;Tang W
种系产生配子，将遗传和表观遗传信息传递给下一代。生殖细胞的维持和配子的发育需要生殖颗粒——保存完好的无膜且富含RNA的细胞器。由于其动态性质及其在种系中的独特表达，胚芽颗粒的组成是难以捉摸的。使用秀丽隐杆线虫胚芽颗粒（称为 P 颗粒）作为模型系统，我们采用基于邻近的标记方法结合质谱法来全面定义其蛋白质成分。这组实验鉴定了 200 多种蛋白质，其中许多含有本质无序区域 (IDR)。基于 RNA 干扰的筛选确定了 P 颗粒组装所必需的因子，特别是 EGGD-1 和 EGGD-2（两种假定的 LOTUS 结构域蛋白）。 Eggd-1 和 Eggd-2 的缺失会导致 P 颗粒与核膜分离、种系萎缩和生育力降低。我们表明，EGGD-1 的 IDR 需要将 EGGD-1 锚定到核周围，而其 LOTUS 结构域需要促进 P 颗粒的核周定位。总而言之，我们的工作扩展了 P 颗粒成分的范围，并为 LOTUS 结构域蛋白在胚芽颗粒组织中的作用提供了新的见解。
The germ line produces gametes that transmit genetic and epigenetic information to the next generation. Maintenance of germ cells and development of gametes require germ granules—well-conserved membraneless and RNA-rich organelles. The composition of germ granules is elusive owing to their dynamic nature and their exclusive expression in the germ line. Using Caenorhabditis elegans germ granule, called P granule, as a model system, we employed a proximity-based labeling method in combination with mass spectrometry to comprehensively define its protein components. This set of experiments identified over 200 proteins, many of which contain intrinsically disordered regions (IDRs). An RNA interference-based screen identified factors that are essential for P granule assembly, notably EGGD-1 and EGGD-2, two putative LOTUS-domain proteins. Loss of eggd-1 and eggd-2 results in separation of P granules from the nuclear envelope, germline atrophy, and reduced fertility. We show that IDRs of EGGD-1 are required to anchor EGGD-1 to the nuclear periphery while its LOTUS domains are required to promote the perinuclear localization of P granules. Taken together, our work expands the repertoire of P granule constituents and provides new insights into the role of LOTUS-domain proteins in germ granule organization.
DOI：
10.2741/3323
发表时间：
2009-01-01
期刊：
Frontiers in bioscience (Landmark edition)
影响因子：
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作者：
Kitagawa R
通讯作者：
Kitagawa R