ES-mediated chimera analysis revealed requirement of DDX6 for NANOS2 localization and function in mouse germ cells
ES-mediated chimera analysis revealed requirement of DDX6 for NANOS2 localization and function in mouse germ cells
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ES介导的嵌合体分析揭示了DDX6对小鼠生殖细胞中NANOS2定位和功能的需求
DOI：
10.1038/s41598-018-36502-0
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发表时间：
2019
影响因子：
4.6
通讯作者：
Saga Yumiko
中科院分区：
文献类型：
--
作者：
Shimada Ryuki;Kiso Makoto;Saga Yumiko
在胚胎雄性生殖细胞中，rna结合蛋白NANOS2将其靶rna招募到加工体（p体），在那里它们被抑制。这一过程是促进雄性型生殖细胞分化所必需的。然而，目前尚不清楚是否所有NANOS2功能都依赖于p体。为了解决这个问题，我们建立了含有生殖细胞特异性诱导Cre和报告基因以及floxedddx6等位基因的ES细胞系。我们通过给含有重组胚胎干细胞衍生的生殖细胞的嵌合胚胎施用他莫昔芬来删除ddx6基因。ddx6缺失的生殖细胞与nanos2缺失的生殖细胞表现出相似和不同的缺陷。这些结果表明，NANOS2的功能是通过p体依赖和p体独立的机制来实现的。RNA-seq分析进一步支持DDX6-null和NANOS2-null生殖细胞之间的表型差异，并表明nanos2介导的基因调控涉及不同的分子级联反应。
In embryonic male germ cells, the RNA-binding protein NANOS2 recruits its target RNAs to processing bodies (P-bodies), where they are repressed. This process is necessary to promote male-type germ cell differentiation. However, it remains unclear whether all NANOS2 functions depend on P-bodies. To address this question, we established ES cell lines containing a germ cell-specific inducible Cre and reporter together with the floxedDdx6allele. We deleted theDdx6gene by administering tamoxifen to chimeric embryos containing germ cells derived from recombinant ES cells. DDX6-null germ cells exhibited both similar and distinct defects from those observed in NANOS2-null germ cells. These results demonstrate that NANOS2 function is carried out via both P-body-dependent and -independent mechanisms. RNA-seq analyses further supported the phenotypic differences between DDX6-null and NANOS2-null germ cells, and indicated distinct molecular cascades involved in NANOS2-mediated gene regulation.