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首页> 外文期刊>Molecular and Cellular Biology >In vivo antisense oligodeoxynucleotide mapping reveals masked regulatory elements in an mRNA dormant in mouse oocytes.
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In vivo antisense oligodeoxynucleotide mapping reveals masked regulatory elements in an mRNA dormant in mouse oocytes.

机译：体内反义寡聚脱氧核苷酸图谱揭示了在小鼠卵母细胞中休眠的mRNA中掩盖的调节元件。

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In mouse oocytes, tissue-type plasminogen activator (tPA) mRNA is under translational control. The newly transcribed mRNA undergoes deadenylation and translational silencing in growing oocytes, while readenylation and translation occur during meiotic maturation. To localize regulatory elements controlling tPA mRNA expression, we identified regions of the endogenous transcript protected from hybridization with injected antisense oligodeoxynucleotides. Most of the targeted sequences in either the 5' untranslated region (5'UTR), coding region, or 3'UTR were accessible to hybridization, as revealed by inhibition of tPA synthesis and by RNase protection. Two protected regions were identified in the 3'UTR of tPA mRNA in primary oocytes: the adenylation control element (ACE) and the AAUAAA polyadenylation signal. These sequences were previously shown to be involved in the translational control of injected reporter transcripts. During the first hour of meiotic maturation, part of the ACE and the AAUAAA hexanucleotide became accessible to hybridization, suggesting a partial unmasking of the 3'UTR of this mRNA before it becomes translationally competent. Our results demonstrate that in vivo antisense oligodeoxynucleotide mapping can reveal the dynamics of regulatory features of a native mRNA in the context of the intact cell. They suggest that specific regions in the 3'UTR of tPA mRNA function as cis-acting masking determinants involved in the silencing of tPA mRNA in primary oocytes.
机译：在小鼠卵母细胞中，组织型纤溶酶原激活物（tPA）mRNA受翻译控制。新转录的mRNA在生长的卵母细胞中经历了腺苷酸化和翻译沉默，而在减数分裂成熟期间发生了腺苷酸化和翻译。为了定位控制tPA mRNA表达的调控元件，我们确定了内源转录物的区域，该区域可防止与注射的反义寡脱氧核苷酸杂交。通过抑制tPA合成和通过RNA酶保护可以发现，5'非翻译区（5'UTR），编码区或3'UTR中的大多数靶向序列均易于杂交。在原代卵母细胞的tPA mRNA的3'UTR中鉴定出两个受保护区域：腺苷酸化控制元件（ACE）和AAUAAA聚腺苷酸化信号。先前已证明这些序列参与注射的报道基因转录的翻译控制。在减数分裂成熟的第一个小时，部分ACE和AAUAAA六核苷酸变得可杂交，这表明该mRNA的3'UTR在其翻译能力之前已部分揭露。我们的结果表明，体内反义寡聚脱氧核苷酸图谱可以揭示完整细胞情况下天然mRNA调控功能的动态。他们认为，tPA mRNA 3'UTR中的特定区域起着顺式作用决定簇的作用，参与了原代卵母细胞中tPA mRNA的沉默。

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