Cargando…
A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance
Imidazopyrazinones (IPYs) are a new class of compounds that target bacterial topoisomerases as a basis for their antibacterial activity. We have characterized the mechanism of these compounds through structural/mechanistic studies showing they bind and stabilize a cleavage complex between DNA gyrase...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5934680/ https://www.ncbi.nlm.nih.gov/pubmed/29538767 http://dx.doi.org/10.1093/nar/gky181 |
_version_ | 1783320160529022976 |
---|---|
author | Germe, Thomas Vörös, Judit Jeannot, Frederic Taillier, Thomas Stavenger, Robert A Bacqué, Eric Maxwell, Anthony Bax, Benjamin D |
author_facet | Germe, Thomas Vörös, Judit Jeannot, Frederic Taillier, Thomas Stavenger, Robert A Bacqué, Eric Maxwell, Anthony Bax, Benjamin D |
author_sort | Germe, Thomas |
collection | PubMed |
description | Imidazopyrazinones (IPYs) are a new class of compounds that target bacterial topoisomerases as a basis for their antibacterial activity. We have characterized the mechanism of these compounds through structural/mechanistic studies showing they bind and stabilize a cleavage complex between DNA gyrase and DNA (‘poisoning’) in an analogous fashion to fluoroquinolones, but without the requirement for the water–metal–ion bridge. Biochemical experiments and structural studies of cleavage complexes of IPYs compared with an uncleaved gyrase–DNA complex, reveal conformational transitions coupled to DNA cleavage at the DNA gate. These involve movement at the GyrA interface and tilting of the TOPRIM domains toward the scissile phosphate coupled to capture of the catalytic metal ion. Our experiments show that these structural transitions are involved generally in poisoning of gyrase by therapeutic compounds and resemble those undergone by the enzyme during its adenosine triphosphate-coupled strand-passage cycle. In addition to resistance mutations affecting residues that directly interact with the compounds, we characterized a mutant (D82N) that inhibits formation of the cleavage complex by the unpoisoned enzyme. The D82N mutant appears to act by stabilizing the binary conformation of DNA gyrase with uncleaved DNA without direct interaction with the compounds. This provides general insight into the resistance mechanisms to antibiotics targeting bacterial type II topoisomerases. |
format | Online Article Text |
id | pubmed-5934680 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-59346802018-05-09 A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance Germe, Thomas Vörös, Judit Jeannot, Frederic Taillier, Thomas Stavenger, Robert A Bacqué, Eric Maxwell, Anthony Bax, Benjamin D Nucleic Acids Res Nucleic Acid Enzymes Imidazopyrazinones (IPYs) are a new class of compounds that target bacterial topoisomerases as a basis for their antibacterial activity. We have characterized the mechanism of these compounds through structural/mechanistic studies showing they bind and stabilize a cleavage complex between DNA gyrase and DNA (‘poisoning’) in an analogous fashion to fluoroquinolones, but without the requirement for the water–metal–ion bridge. Biochemical experiments and structural studies of cleavage complexes of IPYs compared with an uncleaved gyrase–DNA complex, reveal conformational transitions coupled to DNA cleavage at the DNA gate. These involve movement at the GyrA interface and tilting of the TOPRIM domains toward the scissile phosphate coupled to capture of the catalytic metal ion. Our experiments show that these structural transitions are involved generally in poisoning of gyrase by therapeutic compounds and resemble those undergone by the enzyme during its adenosine triphosphate-coupled strand-passage cycle. In addition to resistance mutations affecting residues that directly interact with the compounds, we characterized a mutant (D82N) that inhibits formation of the cleavage complex by the unpoisoned enzyme. The D82N mutant appears to act by stabilizing the binary conformation of DNA gyrase with uncleaved DNA without direct interaction with the compounds. This provides general insight into the resistance mechanisms to antibiotics targeting bacterial type II topoisomerases. Oxford University Press 2018-05-04 2018-03-10 /pmc/articles/PMC5934680/ /pubmed/29538767 http://dx.doi.org/10.1093/nar/gky181 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Nucleic Acid Enzymes Germe, Thomas Vörös, Judit Jeannot, Frederic Taillier, Thomas Stavenger, Robert A Bacqué, Eric Maxwell, Anthony Bax, Benjamin D A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title | A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title_full | A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title_fullStr | A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title_full_unstemmed | A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title_short | A new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in DNA gyrase poisoning and mechanisms of resistance |
title_sort | new class of antibacterials, the imidazopyrazinones, reveal structural transitions involved in dna gyrase poisoning and mechanisms of resistance |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5934680/ https://www.ncbi.nlm.nih.gov/pubmed/29538767 http://dx.doi.org/10.1093/nar/gky181 |
work_keys_str_mv | AT germethomas anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT vorosjudit anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT jeannotfrederic anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT taillierthomas anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT stavengerroberta anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT bacqueeric anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT maxwellanthony anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT baxbenjamind anewclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT germethomas newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT vorosjudit newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT jeannotfrederic newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT taillierthomas newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT stavengerroberta newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT bacqueeric newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT maxwellanthony newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance AT baxbenjamind newclassofantibacterialstheimidazopyrazinonesrevealstructuraltransitionsinvolvedindnagyrasepoisoningandmechanismsofresistance |