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.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alpha-Amanitin / pharmacology
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Amino Acid Motifs
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Amino Acid Sequence
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Catalytic Domain
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Enzyme Inhibitors / pharmacology
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Genetic Complementation Test
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Humans
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Malaria, Falciparum / parasitology
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Plasmodium falciparum / drug effects
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Plasmodium falciparum / genetics*
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Plasmodium falciparum / metabolism
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Protein Subunits / genetics*
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Protein Subunits / metabolism
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Protozoan Proteins / genetics*
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Protozoan Proteins / metabolism
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RNA Polymerase II / genetics*
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RNA Polymerase II / metabolism
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Recombinant Proteins / genetics*
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
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Sequence Homology, Amino Acid
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Small Molecule Libraries
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Transcription, Genetic / drug effects
Substances
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Alpha-Amanitin
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Enzyme Inhibitors
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Protein Subunits
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Protozoan Proteins
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Recombinant Proteins
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Small Molecule Libraries
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RNA Polymerase II