Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei
| Autor: | Nicholas Hasle, Savitha Kalidas, Mehdi Labaied, Margaret A. Phillips, Igor Cestari, Kenneth Stuart, Severine Monnerat, Qiong Li, Marilyn Parsons, Sandesh Regmi |
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| Rok vydání: | 2014 |
| Předmět: |
Trypanosoma brucei brucei
Protozoan Proteins Trypanosoma brucei Microbiology Amino Acyl-tRNA Synthetases chemistry.chemical_compound Cytosol RNA interference Gene expression parasitic diseases Drug Discovery RNA Messenger RNA Small Interfering Molecular Biology Gene chemistry.chemical_classification Gene knockdown biology Aminoacyl tRNA synthetase Drug discovery Cell Cycle General Medicine Articles biology.organism_classification Trypanocidal Agents Mitochondria Enzyme chemistry Biochemistry Gene Expression Regulation Gene Knockdown Techniques Fatty Alcohols Signal Transduction |
| Zdroj: | Eukaryotic cell. 13(4) |
| ISSN: | 1535-9786 |
| Popis: | Human African trypanosomiasis (HAT) is an important public health threat in sub-Saharan Africa. Current drugs are unsatisfactory, and new drugs are being sought. Few validated enzyme targets are available to support drug discovery efforts, so our goal was to obtain essentiality data on genes with proven utility as drug targets. Aminoacyl-tRNA synthetases (aaRSs) are known drug targets for bacterial and fungal pathogens and are required for protein synthesis. Here we survey the essentiality of eight Trypanosoma brucei aaRSs by RNA interference (RNAi) gene expression knockdown, covering an enzyme from each major aaRS class: valyl-tRNA synthetase (ValRS) (class Ia), tryptophanyl-tRNA synthetase (TrpRS-1) (class Ib), arginyl-tRNA synthetase (ArgRS) (class Ic), glutamyl-tRNA synthetase (GluRS) (class 1c), threonyl-tRNA synthetase (ThrRS) (class IIa), asparaginyl-tRNA synthetase (AsnRS) (class IIb), and phenylalanyl-tRNA synthetase (α and β) (PheRS) (class IIc). Knockdown of mRNA encoding these enzymes in T. brucei mammalian stage parasites showed that all were essential for parasite growth and survival in vitro . The reduced expression resulted in growth, morphological, cell cycle, and DNA content abnormalities. ThrRS was characterized in greater detail, showing that the purified recombinant enzyme displayed ThrRS activity and that the protein localized to both the cytosol and mitochondrion. Borrelidin, a known inhibitor of ThrRS, was an inhibitor of T. brucei ThrRS and showed antitrypanosomal activity. The data show that aaRSs are essential for T. brucei survival and are likely to be excellent targets for drug discovery efforts. |
| Databáze: | OpenAIRE |
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