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首页> 外文期刊>Journal of Andrology >Identification of potential regulatory elements in the 5′ and 3′ UTRs of 12 translationally regulated mRNAs in mammalian spermatids by comparative genomics
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Identification of potential regulatory elements in the 5′ and 3′ UTRs of 12 translationally regulated mRNAs in mammalian spermatids by comparative genomics

机译：通过比较基因组学鉴定哺乳动物精子中12种翻译调控mRNA的5'和3'UTR中的潜在调控元件

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摘要

To facilitate identifying translational control elements by studies of mutations in transgenic mice, a database of orthologous 5′ and 3′ ends of 12 messenger RNA (mRNA) species from 13 to 23 mammals that undergo delayed translational activation in spermatids was constructed for the Acev2, Akap3, Akap4v2, Gapdhs, Odf1, Prm1, Prm2, Prm3, Smcp, Spata18, Tnp1, and Tnp2 mRNAs. This database, available here, was searched for conserved sequences in conserved positions and known translational control elements. Numerous potential mRNA-specific elements were identified, including upstream open reading frames, conserved sequences upstream and downstream of the poly(A) signal, and noncanonical and multiple poly(A) signals. RNA electrophoresis mobility shift assays demonstrate that Y-box proteins bind 30 of the 36 permutations of the degenerate Y-box recognition sequence (YRS), [UAC][CA]CA[UC]C[ACU], and this information was used to identify hundreds of YRSs in the untranslated region (UTR) database. Collectively, these findings suggest that the distal ends of both UTRs are particularly well conserved, implying that translation of each mRNA is regulated by mechanisms involving the poly(A) binding protein and the closed loop. In addition, the 5′ flanking regions of all 12 genes have conserved, gene-specific sequences and configurations of elements that resemble the binding site of the testis-specific isoform of cyclic AMP response element modulator, and all 12 genes lack retrogene paralogues, demonstrating the efficacy of mechanisms that limit the proliferation of retroposons in the male germ line. This study illustrates the power of comparative genomics in identifying novel hypothetical regulatory elements for analysis with biochemical and in vivo genetic approaches.
机译：为了方便通过研究转基因小鼠中的突变来识别翻译控制元件，针对Acev2，构建了13种至23种哺乳动物的12个信使RNA（mRNA）直系同源物的5'和3'末端数据库，这些哺乳动物在精子中经历了延迟的翻译激活， Akap3，Akap4v2，Gapdhs，Odf1，Prm1，Prm2，Prm3，Smcp，​​Spata18，Tnp1和Tnp2 mRNA。在此可用的数据库中搜索了保守位置的保守序列和已知的翻译控制元件。确定了许多潜在的mRNA特异元件，包括上游开放阅读框，poly（A）信号上游和下游的保守序列以及非规范和多个poly（A）信号。 RNA电泳迁移率变动分析表明，Y-box蛋白结合了简并Y-box识别序列（YRS），[UAC] [CA] CA [UC] C [ACU]的36个排列中的30个，并用于在非翻译区域（UTR）数据库中识别数百个YRS。总体而言，这些发现表明两个UTR的末端特别保守，这意味着每个mRNA的翻译受涉及poly（A）结合蛋白和闭环的机制调控。此外，所有12个基因的5'侧翼区域均具有保守的，基因特异性的序列和元件构型，类似于环状AMP反应元件调节剂的睾丸特异性同工型的结合位点，并且所有12个基因均缺乏逆转录旁系同源物，表明限制逆转录子在雄性种系中增殖的机制的功效。这项研究说明了比较基因组学在鉴定新颖的假设调控元件以进行生化和体内遗传方法分析中的作用。

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