蛤卵母细胞中隐藏mrna翻译的控制。

Enzyme Pub Date : 1990-01-01 DOI:10.1159/000468751
N Standart, T Hunt
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引用次数: 13

摘要

蛤卵细胞受精后,蛋白质合成模式发生改变。在翻译量增加的mrna中,数量最多的是编码核糖核苷酸还原酶和A型和b型细胞周期蛋白。这些mrna已经被克隆和测序,但它们的序列并没有显示出可以解释掩蔽机制的相似区域。然而,这些mrna在无细胞翻译实验中保持“隐藏”状态,它们的翻译可以通过高盐凝胶过滤激活,这可能会去除抑制蛋白。“竞争性揭膜”试验用于鉴定每个mRNA的蛋白质结合区域。这包括添加反义RNA的短片段,这些片段可以退火到mRNA并取代阻遏物。该实验揭示的核糖核苷酸还原酶和细胞周期蛋白A mrna的揭开区长120-140 nt,位于3'非编码区的中心部分。
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Control of translation of masked mRNAs in clam oocytes.

The pattern of protein synthesis changes soon after fertilization of clam oocytes. The most abundant of the mRNAs whose translation increases at this time encode ribonucleotide reductase and the A- and B-type cyclins. These mRNAs have been cloned and sequenced, yet their sequences do not show regions of similarity that could explain the masking mechanism. However, these mRNAs retain their 'masked' state in cell-free translation assays and their translation can be activated by gel filtration in high salt, which probably removes repressor proteins. A 'competitive unmasking' assay was used to identify the protein-binding regions of each mRNA. This involved adding short segments of antisense RNA that annealed to the mRNA and displaced the repressors. The unmasking regions in ribonucleotide reductase and cyclin A mRNAs revealed by this assay are 120-140 nt long and are located in the central portions of the 3' non-coding regions.

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