Inhibition of translational initiation in the yeast Saccharomyces cerevisiae as a function of the stability and position of hairpin structures in the mRNA leader.

摘要: A new modular in vivo/in vitro expression system was constructed which facilitates studies of the control and regulation gene yeast Saccharomyces cerevisiae. We studied influence stem-loop structures inserted into non-translated leader region upon steady-state levels translation mRNAs bearing cat from bacterial transposon Tn9. mRNA abundance changed relatively little response to alterations sequence structure, whereas clearly inhibited a degree that dependent predicted stability as well position secondary structure. structure with greater than -28 kcal mol-1 stem comprising at least 15 (mainly G/C) base pairs vivo by 98%. approximately -14 mol-1, whose comprised six G/C pairs, 66%. The hairpins were more inhibitory when placed close start codon positioned near 5' end leader. An showing extensive complementarity between trailer regions not only poorly translated but also had level three times higher average for all constructs examined. Translation various cell-free followed qualitatively same pattern results obtained vivo. far less reticulocyte lysate system. Overall, data are likely reflect full spectrum translational activities determined emphasize importance step expression.