ZIF-1 介导的秀丽隐杆线虫生殖系中锌指蛋白的降解。
ZIF-1-mediated degradation of zinc finger proteins in the Caenorhabditis elegans germ line.
机构信息
Department of Cell Biology, NYU Grossman School of Medicine, New York, NY 10016, USA.
Skirball Institute of Biomolecular Medicine, NYU Grossman School of Medicine, New York, NY 10016, USA.
出版信息
Genetics. 2023 Nov 1;225(3). doi: 10.1093/genetics/iyad160.
Rapid and conditional protein depletion is the gold standard genetic tool for deciphering the molecular basis of developmental processes. Previously, we showed that by conditionally expressing the E3 ligase substrate adaptor ZIF-1 in Caenorhabditis elegans somatic cells, proteins tagged with the first CCCH Zn finger 1 (ZF1) domain from the germline regulator PIE-1 degrade rapidly, resulting in loss-of-function phenotypes. The described role of ZIF-1 is to clear PIE-1 and several other CCCH Zn finger proteins from early somatic cells, helping to enrich them in germline precursor cells. Here, we show that proteins tagged with the PIE-1 ZF1 domain are subsequently cleared from primordial germ cells (PGCs) in embryos and from undifferentiated germ cells in larvae and adults by ZIF-1. We harness germline ZIF-1 activity to degrade a ZF1-tagged fusion protein from PGCs and show that its depletion produces phenotypes equivalent to those of a null mutation. Our findings reveal that ZIF-1 transitions from degrading CCCH Zn finger proteins in somatic cells to clearing them from undifferentiated germ cells, and that ZIF-1 activity can be harnessed as a new genetic tool to study the early germline.
快速且有条件的蛋白质耗竭是解析发育过程分子基础的黄金标准遗传工具。此前,我们表明通过在秀丽隐杆线虫体细胞中条件性表达 E3 连接酶底物衔接蛋白 ZIF-1,来自生殖系调控因子 PIE-1 的带有第一个 CCCH Zn 指 1(ZF1)结构域的蛋白质会迅速降解,导致功能丧失表型。所描述的 ZIF-1 作用是从早期体细胞中清除 PIE-1 和其他几个 CCCH Zn 指蛋白,有助于将它们富集到生殖系前体细胞中。在这里,我们表明,带有 PIE-1 ZF1 结构域的蛋白质随后会被 ZIF-1 从胚胎中的原始生殖细胞(PGC)和幼虫及成虫中的未分化生殖细胞中清除。我们利用生殖系 ZIF-1 活性来降解 PGC 中带有 ZF1 标签的融合蛋白,并表明其耗竭会产生与 null 突变相当的表型。我们的发现表明,ZIF-1 从降解体细胞中的 CCCH Zn 指蛋白转变为从未分化的生殖细胞中清除它们,并且 ZIF-1 活性可以作为一种新的遗传工具来研究早期生殖系。