Tindell Samuel J, Boeving Alyssa G, Aebersold Julia, Arkov Alexey L
Department of Biological Sciences, Murray State University, Murray, KY, USA.
Micro/Nano Technology Center, University of Louisville, Louisville, KY, USA.
bioRxiv. 2025 Mar 17:2025.03.13.643173. doi: 10.1101/2025.03.13.643173.
Scaffold proteins play crucial roles in subcellular organization and function. In many organisms, proteins with multiple Tudor domains are required for the assembly of membraneless RNA-protein organelles (germ granules) in germ cells. Tudor domains are protein-protein interaction modules which bind to methylated polypeptides. Tudor protein contains eleven Tudor domains, which is the highest number known in a single protein. The role of each of these domains in germ cell formation has not been systematically tested and it is not clear if some domains are functionally redundant. Using CRISPR methodology, we generated mutations in several uncharacterized Tudor domains and showed that they all caused defects in germ cell formation. Mutations in individual domains affected Tudor protein differently causing reduction in protein levels, defects in subcellular localization and in the assembly of germ granules. Our data suggest that multiple domains of Tudor protein are all needed for efficient germ cell formation highlighting the rational for keeping many Tudor domains in protein scaffolds of biomolecular condensates in and other organisms.
支架蛋白在亚细胞组织和功能中发挥着关键作用。在许多生物体中，生殖细胞中无膜RNA-蛋白质细胞器（生殖颗粒）的组装需要具有多个 Tudor 结构域的蛋白质。Tudor 结构域是与甲基化多肽结合的蛋白质-蛋白质相互作用模块。Tudor 蛋白包含 11 个 Tudor 结构域，这是单个蛋白质中已知数量最多的。这些结构域在生殖细胞形成中的作用尚未得到系统测试，目前尚不清楚某些结构域是否在功能上冗余。利用CRISPR方法，我们在几个未表征的 Tudor 结构域中产生了突变，并表明它们都导致了生殖细胞形成缺陷。单个结构域的突变对 Tudor 蛋白的影响不同，导致蛋白质水平降低、亚细胞定位缺陷以及生殖颗粒组装缺陷。我们的数据表明，Tudor 蛋白的多个结构域对于高效的生殖细胞形成都是必需的，这突出了在[具体生物]和其他生物体的生物分子凝聚物的蛋白质支架中保留多个 Tudor 结构域的合理性。
