A single amino acid substitution (H451Y) in Leishmania calcium-dependent kinase SCAMK confers high tolerance and resistance to antimony
| Autor: | Baptiste Vergnes, Yvon Sterkers, Lauriane Sollelis, Cédric Mariac, Anne-Laure Bañuls, Miléna Du Manoir, Jose-Juan Lopez-Rubio, Elodie Gazanion |
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| Přispěvatelé: | Du gène à l'écosystème (MIVEGEC-GeneSys), Pathogènes, Environnement, Santé Humaine (EPATH), Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Diversité, adaptation, développement des plantes (UMR DIADE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Biologie, Génétique et Pathologie des Pathogènes Eucaryotes (MIVEGEC-BioGEPPE), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Institut de Recherche pour le Développement (IRD [France-Sud])-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Antimony
0301 basic medicine Microbiology (medical) 030106 microbiology Mutant Antiprotozoal Agents Drug Resistance Protozoan Proteins Trypanothione Drug resistance 03 medical and health sciences chemistry.chemical_compound Parasitic Sensitivity Tests Pharmacology (medical) [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Mode of action Gene Editing Leishmania Pharmacology biology Chemistry Resistance mutation biology.organism_classification Nicotinamidase 3. Good health 030104 developmental biology Infectious Diseases Amino Acid Substitution Biochemistry Mutation [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology Calcium NAD+ kinase CRISPR-Cas Systems |
| Zdroj: | Journal of Antimicrobial Chemotherapy Journal of Antimicrobial Chemotherapy, 2019, 74 (11), pp.3231-3239. ⟨10.1093/jac/dkz334⟩ Journal of Antimicrobial Chemotherapy, Oxford University Press (OUP), 2019, 74 (11), pp.3231-3239. ⟨10.1093/jac/dkz334⟩ |
| ISSN: | 0305-7453 1460-2091 |
| DOI: | 10.1093/jac/dkz334⟩ |
| Popis: | Background For almost a century, antimonials have remained the first-line drugs for the treatment of leishmaniasis. However, little is known about their mode of action and clinical resistance mechanisms. Objectives We have previously shown that Leishmania nicotinamidase (PNC1) is an essential enzyme for parasite NAD+ homeostasis and virulence in vivo. Here, we found that parasites lacking the pnc1 gene (Δpnc1) are hypersusceptible to the active form of antimony (SbIII) and used these mutant parasites to better understand antimony’s mode of action and the mechanisms leading to resistance. Methods SbIII-resistant WT and Δpnc1 parasites were selected in vitro by a stepwise selection method. NAD(H)/NADP(H) dosages and quantitative RT–PCR experiments were performed to explain the susceptibility differences observed between strains. WGS and a marker-free CRISPR/Cas9 base-editing approach were used to identify and validate the role of a new resistance mutation. Results NAD+-depleted Δpnc1 parasites were highly susceptible to SbIII and this phenotype could be rescued by NAD+ precursor or trypanothione precursor supplementation. Δpnc1 parasites could become resistant to SbIII by an unknown mechanism. WGS revealed a unique amino acid substitution (H451Y) in an EF-hand domain of an orphan calcium-dependent kinase, recently named SCAMK. When introduced into a WT reference strain by base editing, the H451Y mutation allowed Leishmania parasites to survive at extreme concentrations of SbIII, potentiating the rapid emergence of resistant parasites. Conclusions These results establish that Leishmania SCAMK is a new central hub of antimony’s mode of action and resistance development, and uncover the importance of drug tolerance mutations in the evolution of parasite drug resistance. |
| Databáze: | OpenAIRE |
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