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首页> 外文期刊>Biochimica et Biophysica Acta. Gene Regulatory Mechanisms >Mouse Dazl and its novel splice variant functions in translational repression of target mRNAs in embryonic stem cells
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Mouse Dazl and its novel splice variant functions in translational repression of target mRNAs in embryonic stem cells

机译：小鼠Dazl及其新型剪接变体在胚胎干细胞中靶标mRNA的翻译抑制中起作用

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摘要

Dazl (deleted in azoospermia-like) is an RNA binding protein that is important for germ cell differentiation in vertebrates. In the present study, we report the identification of a novel Dazl isoform (Dazl_δ8) that results from alternative splicing of exon8 of mouse Dazl. We observed the expression of Dazl_δ8 in various pluripotent cell types, but not in somatic cells. Furthermore, the Dazl_δ8 splice variant was expressed along with the full-length isoform of Dazl (Dazl_FL) throughout male germ-cell development and in the ovary. Sub-cellular localization studies of Dazl_δ8 revealed a diffused cytoplasmic and large granular pattern, which is similar to the localization patterns of Dazl_FL protein. In contrast to the well documented translation stimulation function in germ cells, overexpression and downregulation studies of Dazl isoforms (Dazl_FL and Dazl_δ8) revealed a role for Dazl in the negative translational regulation of Mvh, a known target of Dazl, as well as Oct3/4 and Sox2 in embryonic stem cells (ESCs). In line with these observations, a luciferase reporter assay with the 3'UTRs of Oct3/4 and Mvh confirmed the translational repressive role of Dazl isoforms in ESCs but not in germ cells derived cell line GC-1. Further, we identified several putative target mRNAs of Dazl_FL and Dazl_δ8 in ESCs through RNA-binding immunoprecipitation followed by whole genome transcriptome analysis. Collectively, our results show a translation repression function of Dazl in pluripotent stem cells.
机译：Dazl（无精症样缺失）是一种RNA结合蛋白，对脊椎动物生殖细胞的分化很重要。在本研究中，我们报告了一种新型Dazl同工型（Dazl_δ8）的鉴定，该鉴定是由小鼠Dazl的exon8的可变剪接产生的。我们观察到Dazl_δ8在各种多能细胞类型中的表达，但在体细胞中却没有。此外，在整个男性生殖细胞发育过程中和在卵巢中，Dazl_δ8剪接变体与Dazl的全长同工型（Dazl_FL）一起表达。 Dazl_δ8的亚细胞定位研究揭示了弥散的细胞质和大颗粒模式，类似于Dazl_FL蛋白的定位模式。与记录在生殖细胞中的翻译刺激功能相反，对Dazl亚型（Dazl_FL和Dazl_δ8）的过度表达和下调研究揭示了Dazl在Maz的负翻译调控中的作用，Mvh是Dazl的已知靶标，以及Oct3 / 4和胚胎干细胞（ESC）中的Sox2。与这些观察结果一致，具有Oct3 / 4和Mvh的3'UTR的荧光素酶报告基因测定证实了Dazl同工型在ESC中但在生殖细胞衍生的细胞系GC-1中没有翻译抑制作用。此外，我们通过RNA结合免疫沉淀，然后进行全基因组转录组分析，确定了ESC中Dazl_FL和Dazl_δ8的几种推定靶标mRNA。总的来说，我们的结果表明Dazl在多能干细胞中具有翻译抑制功能。

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