Mezl V A, Clark J, Garber P M
Biochem Cell Biol. 1986 Jun;64(6):504-8. doi: 10.1139/o86-070.
A recent study has reported that the heating of a population of guanidinium-extracted mRNAs prior to translation causes a selective increase in the translation of certain mRNAs. To determine if this phenomenon is a general property of mRNAs, we carried out a comparison of the translation products obtained when phenol-extracted rat heart and mammary gland RNAs, rabbit reticulocyte membrane RNAs, and trout liver RNAs were translated in the reticulocyte translation system, with and without a prior heat treatment. Our results show that no selective increase in the translation of mRNAs was observed for any of these samples. Among the 14 RNA preparations examined, one total mammary RNA preparation did display a twofold increase in the translation of all mRNAs after heat treatment. It is shown that the heat enhancement of translational activity observed for this sample was due to the reversible formation of intermolecular aggregates with a contaminant that can be removed by chromatography on oligo(dT)-cellulose. Since heat treatment did not selectively enhance the activity of any mRNA in these samples, our results show that the current practice of translating phenol-extracted RNAs without a prior heat treatment should be satisfactory for the translation of most mRNA populations.
最近的一项研究报告称,在翻译前对胍盐提取的mRNA群体进行加热会导致某些mRNA的翻译选择性增加。为了确定这种现象是否是mRNA的普遍特性,我们比较了在有或没有预先热处理的情况下,将苯酚提取的大鼠心脏和乳腺RNA、兔网织红细胞膜RNA以及鳟鱼肝脏RNA在网织红细胞翻译系统中进行翻译时获得的翻译产物。我们的结果表明,这些样品中均未观察到mRNA翻译的选择性增加。在所检测的14种RNA制剂中,一种乳腺总RNA制剂在热处理后所有mRNA的翻译确实增加了两倍。结果表明,该样品中观察到的翻译活性的热增强是由于与一种污染物形成了可逆的分子间聚集体,这种污染物可以通过寡聚(dT)-纤维素柱层析去除。由于热处理并未选择性增强这些样品中任何mRNA的活性,我们的结果表明,目前在没有预先热处理的情况下翻译苯酚提取的RNA的做法对于大多数mRNA群体的翻译应该是令人满意的。