Asselbergs F A, Peters W H, van Venrooij W J, Bloemendal H
Biochim Biophys Acta. 1978 Oct 24;520(3):577-87. doi: 10.1016/0005-2787(78)90143-0.
The nuclease treated reticulocyte lysate forms a highly efficient and completely mRNA-dependent cell-free system. In this system the functioning of the cap on eukaryotic mRNAs was explored by blocking cap recognition with cap analogues. Translation of capped mRNAs was severely inhibited, while translation of uncapped mRNAs was unaffected. It is concluded that this cell-free system can be used for screening cap dependence in the translation of specific mRNAs, like calf lens mRNAs. At 1.2 mM m7G5'p, 0.16 mM m7G5'pp or 0.16 m7G5'ppp5'G, translation of all lens mRNAs was totally inhibited. At lower concentrations the sensitivity to cap analogues was different for the various species of lens crystallin messenger. gamma-Crystallin mRNA showed relatively the lowest response. The translation of added polyribosomes was also inhibited by the cap analogue. It is concluded that translation of all crystallin messengers is cap-dependent.
经核酸酶处理的网织红细胞裂解物形成了一个高效且完全依赖mRNA的无细胞体系。在该体系中，通过用帽类似物阻断帽识别来探究真核mRNA上帽的功能。加帽mRNA的翻译受到严重抑制，而未加帽mRNA的翻译则不受影响。得出的结论是，这个无细胞体系可用于筛选特定mRNA（如小牛晶状体mRNA）翻译中的帽依赖性。在1.2 mM m7G5'p、0.16 mM m7G5'pp或0.16 m7G5'ppp5'G时，所有晶状体mRNA的翻译都被完全抑制。在较低浓度下，各种晶状体晶状体蛋白信使对帽类似物的敏感性不同。γ-晶状体蛋白mRNA的反应相对最低。添加的多核糖体的翻译也受到帽类似物的抑制。得出的结论是，所有晶状体蛋白信使的翻译都依赖帽。
