Generation and functional characterization of a conditional Pumilio2 null allele.
Generation and functional characterization of a conditional Pumilio2 null allele.
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DOI：
10.7555/jbr.32.20170117
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发表时间：
2018-11-20
影响因子：
2.3
通讯作者：
Yujun Xu E
中科院分区：
文献类型：
--
作者：
Lin K;Zhang S;Chen J;Yang D;Zhu MY;Yujun Xu E
高度保守的RNA结合蛋白PUF （Pumilio/FBF）家族存在于从酵母到哺乳动物的真核生物中，在发育、生育和神经系统中起着关键作用。然而，哺乳动物PUF家族成员的功能仍未得到充分研究。我们之前的研究报道了Pum2基因诱捕突变导致睾丸变小，但不影响生育能力和生存能力。虽然基因陷阱突变破坏了pumm蛋白- pumm - hd的关键功能域（Pumilio同源结构域），但仍然产生了含有部分Pum2的嵌合Pum2-β-geo蛋白，这就提出了这样的嵌合蛋白是否提供任何残留功能或有助于生殖表型的问题。在这里，我们报告了一个条件PUM2等位基因的产生，当敲除时，不会产生残留的PUM2，因此是一个完全丧失功能的等位基因。我们还发现突变体的雄性生育能力和生存能力虽然小但显著降低，这表明雄性生育能力和生存能力需要PUM2。
The highly conserved RNA binding protein PUF (Pumilio/FBF) family is present throughout eukaryotes from yeast to mammals, with critical roles in development, fertility and the nervous system. However, the function of the mammalian PUF family members remains underexplored. Our previous study reported that a gene-trap mutation of Pum2 results in a smaller testis but does not impact fertility and viability. Although the gene-trap mutation disrupted the key functional domain of PUM protein–PUM-HD (Pumilio homology domain), but still produced a chimeric Pum2-β-geo protein containing part of PUM2, raising a question if such a chimeric protein may provide any residual function or contribute to the reproductive phenotype. Here, we report the generation of a conditional PUM2 allele, when knocked out, producing no residual PUM2 and hence a complete loss-of-function allele. We also uncovered small but significant reduction of male fertility and viability in the mutants, suggesting requirement of PUM2 for male fertility and viability.