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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove
2023
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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. |
format | Online Article Text |
id | pubmed-10460174 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Dove |
record_format | MEDLINE/PubMed |
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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