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Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica

PURPOSE: Elizabethkingia meningoseptica (EM) is a multi-drug-resistant bacterium of global concern for its role in nosocomial infection and is generally resistant to aminoglycoside antibiotics. In the whole genome of an EM strain (FMS-007), an aminoglycoside-6-adenyl transferase gene (ant(6)(FMS-007...

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Autores principales: Zhang, Shaoxing, Zhang, Yuxin, Liu, Ruijie, Yuan, Shuying, Chen, Yanwen, Li, Wenjie, Lu, Xinrong, Tong, Yongliang, Hou, Linlin, Chen, Li, Sun, Guiqin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10460174/
https://www.ncbi.nlm.nih.gov/pubmed/37638067
http://dx.doi.org/10.2147/IDR.S423418
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author Zhang, Shaoxing
Zhang, Yuxin
Liu, Ruijie
Yuan, Shuying
Chen, Yanwen
Li, Wenjie
Lu, Xinrong
Tong, Yongliang
Hou, Linlin
Chen, Li
Sun, Guiqin
author_facet Zhang, Shaoxing
Zhang, Yuxin
Liu, Ruijie
Yuan, Shuying
Chen, Yanwen
Li, Wenjie
Lu, Xinrong
Tong, Yongliang
Hou, Linlin
Chen, Li
Sun, Guiqin
author_sort Zhang, Shaoxing
collection PubMed
description PURPOSE: Elizabethkingia meningoseptica (EM) is a multi-drug-resistant bacterium of global concern for its role in nosocomial infection and is generally resistant to aminoglycoside antibiotics. In the whole genome of an EM strain (FMS-007), an aminoglycoside-6-adenyl transferase gene (ant(6)(FMS-007)) was predicted. This study aimed to characterize the biochemical function of ANT(6)(FMS-007) and analyze the relationship between genotype and phenotype of ant(6) in clinical EM isolates, so as to provide evidence for clinical precision drug use. This study could establish a method for the verification of known or unknown functionally resistant genes. METHODS: A total of 42 EM clinical isolates were collected from clinical departments during 2015–2023. The phenotype of aminoglycoside antibiotics was analyzed by broth microdilution (BMD) and Kirby-Bauer (K-B) methods. The whole-length ant(6) from EM clinical isolates was analyzed by polymerase chain reaction (PCR) and sequencing. The biochemical function of predictive ANT(6)(FMS-007) from the FMS-007 whole genome was identified by 3D plate experiment and mass spectrometry analysis. Candidate active sites were predicted by multi-species sequence alignment and molecular docking, and other important sites were identified in the comparison of ant(6) genotypes and phenotypes of EM clinical isolates. Drug susceptibility test was used to verify the function of these sites. RESULTS: The predictive ANT(6)(FMS-007) protein could inactivate STR by modifying STR with ATP to form STR-AMP. Four active sites (Asp-38, Asp-42, Lys-95, and Lys-213) of ANT(6)(FMS-007) were identified. Thirty-one EM clinical isolates (74%) carried the ant(6) gene. Eight EM clinical isolates containing the ant(6) gene had MIC values (<=32μg/mL) lower by at least 16-fold than FMS-007 (512μg/mL) for STR, and N59H and K204Q were the common mutations in the ant(6) gene. CONCLUSION: This assay verified the biochemical function of the predictive gene ant(6)(FMS-007) and could provide an alternative method to study resistant gene function in multi-drug-resistant bacteria. The inconsistency between genotype and phenotype of resistant genes indicated that the combination of resistance gene detection and functional analysis could better provide precision medicine for clinical use.
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spelling pubmed-104601742023-08-27 Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica Zhang, Shaoxing Zhang, Yuxin Liu, Ruijie Yuan, Shuying Chen, Yanwen Li, Wenjie Lu, Xinrong Tong, Yongliang Hou, Linlin Chen, Li Sun, Guiqin Infect Drug Resist Original Research PURPOSE: Elizabethkingia meningoseptica (EM) is a multi-drug-resistant bacterium of global concern for its role in nosocomial infection and is generally resistant to aminoglycoside antibiotics. In the whole genome of an EM strain (FMS-007), an aminoglycoside-6-adenyl transferase gene (ant(6)(FMS-007)) was predicted. This study aimed to characterize the biochemical function of ANT(6)(FMS-007) and analyze the relationship between genotype and phenotype of ant(6) in clinical EM isolates, so as to provide evidence for clinical precision drug use. This study could establish a method for the verification of known or unknown functionally resistant genes. METHODS: A total of 42 EM clinical isolates were collected from clinical departments during 2015–2023. The phenotype of aminoglycoside antibiotics was analyzed by broth microdilution (BMD) and Kirby-Bauer (K-B) methods. The whole-length ant(6) from EM clinical isolates was analyzed by polymerase chain reaction (PCR) and sequencing. The biochemical function of predictive ANT(6)(FMS-007) from the FMS-007 whole genome was identified by 3D plate experiment and mass spectrometry analysis. Candidate active sites were predicted by multi-species sequence alignment and molecular docking, and other important sites were identified in the comparison of ant(6) genotypes and phenotypes of EM clinical isolates. Drug susceptibility test was used to verify the function of these sites. RESULTS: The predictive ANT(6)(FMS-007) protein could inactivate STR by modifying STR with ATP to form STR-AMP. Four active sites (Asp-38, Asp-42, Lys-95, and Lys-213) of ANT(6)(FMS-007) were identified. Thirty-one EM clinical isolates (74%) carried the ant(6) gene. Eight EM clinical isolates containing the ant(6) gene had MIC values (<=32μg/mL) lower by at least 16-fold than FMS-007 (512μg/mL) for STR, and N59H and K204Q were the common mutations in the ant(6) gene. CONCLUSION: This assay verified the biochemical function of the predictive gene ant(6)(FMS-007) and could provide an alternative method to study resistant gene function in multi-drug-resistant bacteria. The inconsistency between genotype and phenotype of resistant genes indicated that the combination of resistance gene detection and functional analysis could better provide precision medicine for clinical use. Dove 2023-08-22 /pmc/articles/PMC10460174/ /pubmed/37638067 http://dx.doi.org/10.2147/IDR.S423418 Text en © 2023 Zhang et al. https://creativecommons.org/licenses/by-nc/3.0/This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/ (https://creativecommons.org/licenses/by-nc/3.0/) ). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).
spellingShingle Original Research
Zhang, Shaoxing
Zhang, Yuxin
Liu, Ruijie
Yuan, Shuying
Chen, Yanwen
Li, Wenjie
Lu, Xinrong
Tong, Yongliang
Hou, Linlin
Chen, Li
Sun, Guiqin
Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title_full Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title_fullStr Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title_full_unstemmed Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title_short Characterization and Molecular Mechanism of Aminoglycoside-6-Adenyl Transferase Associated with Aminoglycoside Resistance from Elizabethkingia meningoseptica
title_sort characterization and molecular mechanism of aminoglycoside-6-adenyl transferase associated with aminoglycoside resistance from elizabethkingia meningoseptica
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10460174/
https://www.ncbi.nlm.nih.gov/pubmed/37638067
http://dx.doi.org/10.2147/IDR.S423418
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