title: A genetic analysis of Plasmodium falciparum RNA polymerase II subunits in yeast.
abstract: 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.
authors.affiliation: Université de Strasbourg, Institut de Recherche de l'Ecole Supérieure de Biotechnologie de Strasbourg, CNRS-UMR7242, Ecole Supérieure de Biotechnologie de Strasbourg, BP10413, 67412 Strasbourg Illkirch Cedex, France.
authors.forename: Adonis
authors.initials: A
authors.lastname: Hazoume
authors.forename: Kambiz
authors.initials: K
authors.lastname: Naderi
authors.forename: Ermanno
authors.initials: E
authors.lastname: Candolfi
authors.forename: Claude
authors.initials: C
authors.lastname: Kedinger
authors.forename: Bruno
authors.initials: B
authors.lastname: Chatton
authors.forename: Marc
authors.initials: M
authors.lastname: Vigneron
mesh_terms: D053959:Alpha-Amanitin / Q000494:pharmacology; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005816:Genetic Complementation Test; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics* / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D054852:Small Molecule Libraries; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D053959:Alpha-Amanitin; D004791:Enzyme Inhibitors; D021122:Protein Subunits; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D012319:RNA Polymerase II