A genetic analysis of Plasmodium falciparum RNA polymerase II subunits in yeast

RNA polymerase II is an essential nuclear multi subunit enzyme that transcribes nearly the whole genome. Its inhibition by the alpha-amanitin toxin leads to cell death. The enzyme of Plasmodium falciparum remains poorly characterized. Using a complementation assay in yeast as a genetic test, we demonstrate that five Plasmodium putative RNA polymerase subunits are indeed functional in vivo. The active site of this enzyme is built from the two largest subunits. Using site directed mutagenesis we were able to modify the active site of the yeast RNA polymerase II so as to introduce Plasmodium or human structural motifs. The resulting strains allow the screening of chemical libraries for potential specific inhibitors.

Using a complementation test in budding yeast, we genetically identified five subunits of the Plasmodium falciparum RNA polymerase II.Figure optionsDownload high-quality image (313 K)Download as PowerPoint slideResearch highlights▶ Test of protozoan RNA polymerase II subunits in yeast complementation assays. ▶ Genetic identification of five Plasmodium falciparum RNA polymerase II subunits. ▶ RNA polymerase II active site modifications.