The steroid derivative 6-aminocholestanol inhibits the DEAD-box helicase eIF4A (LieIF4A) from the Trypanosomatid parasite Leishmania by perturbing the RNA and ATP binding sites
| Autor: | N. Kyle Tanner, Mourad Barhoumi, Emna Harigua-Souiai, Arnaud Blondel, Josette Banroques, Yosser Zina Abdelkrim, Ikram Guizani |
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| Přispěvatelé: | Expression Génétique Microbienne (EGM (UMR_8261 / FRE_3630)), Institut de biologie physico-chimique (IBPC (FR_550)), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP), Faculté des Sciences de Bizerte [Université de Carthage], Université de Carthage - University of Carthage, Bioinformatique structurale - Structural Bioinformatics, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the Centre National de la Recherche Scientifique, France, by the HelicaRN [2010 BLAN 1503 01] and HeliDEAD grants [ANR-13-BSV8-0009-01] from the Agence Nationale de la Recherche, France, and by the Initiative d'Excellence program from the French State [Grant DYNAMO, ANR-11-LABX-0011-01] to NKT. This work received financial support from the Pasteur Institute Transversal Research Program (grant PTR426), France, and partially from the Ministry of Higher Education and Research in Tunisia (LR11IPT04 & LR16IPT04) to IG. EHS received support from the UNESCO-L'Oréal, 'For Women in Science,' international fellowship and the Institut Pasteur International Network (Calmette and Yersin programme)., We thank Patrick Linder, Centre Médical Universitaire, Geneva, Switzerland, for the eIF4AIMus clone. The 6-aminocholestanol was a gift from Dr. Leila el Kihel, Centre d'Etudes et de Recherche sur le Médicament de Normandie (CERMN), UFR des Sciences Pharmaceutiques, Université de Caen de Basse-Normandie, France, for the minimal costs of shipping fees. We thank Damien Monet for his assistance in the cavity generation and analyses., ANR-13-BSV8-0009,HeliDEAD,Les ARN hélicases à boite DEAD: quels sont leurs rôles, leurs partenaires, leurs substrats ARN et comment elles fonctionnent.(2013), ANR-11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences de Bizerte, Bioinformatique structurale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), ANR-11-LABX-0011/11-LABX-0011,DYNAMO,Dynamique des membranes transductrices d'énergie : biogénèse et organisation supramoléculaire.(2011), Institut de biologie physico-chimique (IBPC) |
| Rok vydání: | 2018 |
| Předmět: |
Hippuristanol
0301 basic medicine Antifungal Agents MESH: Sequence Homology Amino Acid ATPase Protozoan Proteins Gene Expression MESH: Protein Structure Secondary MESH: Amino Acid Sequence Protein Structure Secondary MESH: Eukaryotic Initiation Factor-4A MESH: Recombinant Proteins Mice chemistry.chemical_compound Adenosine Triphosphate MESH: Cholesterol MESH: Genetic Vectors MESH: Adenosine Triphosphate MESH: Animals MESH: Trypanocidal Agents Cloning Molecular Leishmania infantum MESH: Protozoan Proteins Conserved Sequence Ergosterol MESH: Conserved Sequence MESH: Kinetics [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] MESH: Escherichia coli Trypanocidal Agents RNA Helicase A Recombinant Proteins MESH: Leishmania infantum [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Cholesterol Biochemistry Translation-initiation factor MESH: Drug Repositioning Protein Binding RNA helicase MESH: Gene Expression Genetic Vectors MESH: Sequence Alignment MESH: Binding Competitive Biology Binding Competitive Drug design 03 medical and health sciences Escherichia coli Animals Humans MESH: Protein Binding [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Protein Interaction Domains and Motifs MESH: Cloning Molecular Amino Acid Sequence [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Binding site MESH: Mice Molecular Biology MESH: Protein Interaction Domains and Motifs Binding Sites MESH: Humans Sequence Homology Amino Acid MESH: RNA Helminth 030102 biochemistry & molecular biology Drug Repositioning RNA Helicase MESH: Antifungal Agents Kinetics 030104 developmental biology MESH: Binding Sites chemistry eIF4A Eukaryotic Initiation Factor-4A Nucleic acid biology.protein Parasitology RNA Helminth Sequence Alignment |
| Zdroj: | Molecular and Biochemical Parasitology Molecular and Biochemical Parasitology, 2018, 226, pp.9-19. ⟨10.1016/j.molbiopara.2018.10.001⟩ Molecular and Biochemical Parasitology, Elsevier, 2018, 226, pp.9-19. ⟨10.1016/j.molbiopara.2018.10.001⟩ |
| ISSN: | 0166-6851 |
| DOI: | 10.1016/j.molbiopara.2018.10.001 |
| Popis: | International audience; The antifungal agent 6-aminocholestanol targets the production of ergosterol, which is the principle sterol in many fungi and protozoans; ergosterol serves many of the same roles as cholesterol in animals. We found that it also is an effective inhibitor of the translation-initiation factor eIF4AI from mouse (eIF4AIMus) and the Trypanosomatid parasite Leishmania (LieIF4A). The eIF4A proteins belong to the DEAD-box family of RNA helicases, which are ATP-dependent RNA-binding proteins and RNA-dependent ATPases. DEAD-box proteins contain a commonly-shared core structure consisting of two linked domains with structural homology to that of recombinant protein A (RecA) and that contain conserved motifs that are involved in RNA and ATP binding, and in the enzymatic activity. The compound inhibits both the ATPase and helicase activities by perturbing ATP and RNA binding, and it is capable of binding other proteins containing nucleic acid-binding sites as well. We undertook kinetic analyses and found that the Leishmania LieIF4A protein binds 6-aminocholestanol with a higher apparent affinity than for ATP, although multiple binding sites were probably involved. Competition experiments with the individual RecA-like domains indicate that the primary binding sites are on RecA-like domain 1, and they include a cavity that we previously identified by molecular modeling of LieIF4A that involve conserved RNA-binding motifs. The compound affects the mammalian and Leishmania proteins differently, which indicates the binding sites and affinities are not the same. Thus, it is possible to develop drugs that target DEAD-box proteins from different organisms even when they are implicated in the same biological process. |
| Databáze: | OpenAIRE |
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