Inhibition of translation of lens mRNAs in a messenger dependent reticulocyte lysate by cap analogues.

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.