Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death

ABSTRACT Trimethoprim-sulfamethoxazole (SXT) is commonly used to treat diverse Staphylococcus aureus infections, including those associated with cystic fibrosis (CF) pulmonary disease. Studies with Escherichia coli found that SXT impairs tetrahydrofolate production, leading to DNA damage, stress res...

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Main Authors: Lauren J. Gonsalves, Allyson Tran, Tessa Gardiner, Tiia Freeman, Angshita Dutta, Carson J. Miller, Sharon McNamara, Adam Waalkes, Dustin R. Long, Stephen J. Salipante, Lucas R. Hoffman, Daniel J. Wolter
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Language:English
Published: American Society for Microbiology 2024-11-01
Series:mBio
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Online Access:https://journals.asm.org/doi/10.1128/mbio.01634-24
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author Lauren J. Gonsalves
Allyson Tran
Tessa Gardiner
Tiia Freeman
Angshita Dutta
Carson J. Miller
Sharon McNamara
Adam Waalkes
Dustin R. Long
Stephen J. Salipante
Lucas R. Hoffman
Daniel J. Wolter
author_facet Lauren J. Gonsalves
Allyson Tran
Tessa Gardiner
Tiia Freeman
Angshita Dutta
Carson J. Miller
Sharon McNamara
Adam Waalkes
Dustin R. Long
Stephen J. Salipante
Lucas R. Hoffman
Daniel J. Wolter
author_sort Lauren J. Gonsalves
collection DOAJ
description ABSTRACT Trimethoprim-sulfamethoxazole (SXT) is commonly used to treat diverse Staphylococcus aureus infections, including those associated with cystic fibrosis (CF) pulmonary disease. Studies with Escherichia coli found that SXT impairs tetrahydrofolate production, leading to DNA damage, stress response induction, and accumulation of reactive oxygen species (ROS) in a process known as thymineless death (TLD). TLD survival can occur through the uptake of exogenous thymidine, countering the effects of SXT; however, a growing body of research has implicated central metabolism as another potentially important determinant of bacterial survival of SXT and other antibiotics. Here, we conducted studies to better understand the mechanisms of TLD survival in S. aureus. We found that thymidine abundances in CF sputum were insufficient to prevent TLD of S. aureus, highlighting the importance of alternative survival mechanisms in vivo. In S. aureus cultured in vitro with SXT and low thymidine, we frequently identified adaptive mutations in genes encoding carbohydrate, nucleotide, and amino acid metabolism, supporting reduced metabolism as a common survival mechanism. Although intracellular ROS levels rose with SXT treatment in vitro, survival was not improved in the presence of ROS scavengers, unlike in E. coli. SXT challenge induced the SOS response, which was alleviated by added thymidine. Finally, an inactivating mutation in the phosphotransferase gene ptsI conferred both limitation in cellular ATP and improved survival against TLD. Collectively, these results suggest that alterations in core metabolic functions, particularly those that reduce ATP levels, predominantly confer S. aureus survival and persistence during SXT treatment, potentially identifying novel targets for co-treatment.IMPORTANCEStaphylococcus aureus is a ubiquitous organism and one of the leading causes of human infections, many of which are difficult to treat due to persistence, antibiotic resistance, or antibiotic tolerance. As our arsenal of effective antibiotics dwindles, the need for improved treatments becomes increasingly urgent, necessitating a better understanding of the precise mechanisms by which pathogens evade our most critical antimicrobial agents. Here, we report a systematic characterization of the mechanisms of S. aureus survival to treatment with the first-line antistaphylococcal antibiotic trimethoprim-sulfamethoxazole, identifying pathways and candidate targets for enhancing the efficacy of available antimicrobial agents.
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spelling doaj-art-448c71145f6a4c8bb0d64a890bcaa0672024-11-13T14:03:28ZengAmerican Society for MicrobiologymBio2150-75112024-11-01151110.1128/mbio.01634-24Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless deathLauren J. Gonsalves0Allyson Tran1Tessa Gardiner2Tiia Freeman3Angshita Dutta4Carson J. Miller5Sharon McNamara6Adam Waalkes7Dustin R. Long8Stephen J. Salipante9Lucas R. Hoffman10Daniel J. Wolter11Department of Microbiology, University of Washington, Seattle, Washington, USADivision of Pulmonary and Sleep Medicine, Department of Pediatrics, University of Washington, Seattle, Washington, USADepartment of Microbiology, University of Washington, Seattle, Washington, USADivision of Pulmonary and Sleep Medicine, Department of Pediatrics, University of Washington, Seattle, Washington, USADivision of Pulmonary and Sleep Medicine, Department of Pediatrics, University of Washington, Seattle, Washington, USADepartment of Microbiology, University of Washington, Seattle, Washington, USAPulmonary Division, Seattle Children’s Hospital, Seattle, Washington, USADepartment of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USADivision of Critical Care Medicine, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, Washington, USADepartment of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USADepartment of Microbiology, University of Washington, Seattle, Washington, USADivision of Pulmonary and Sleep Medicine, Department of Pediatrics, University of Washington, Seattle, Washington, USAABSTRACT Trimethoprim-sulfamethoxazole (SXT) is commonly used to treat diverse Staphylococcus aureus infections, including those associated with cystic fibrosis (CF) pulmonary disease. Studies with Escherichia coli found that SXT impairs tetrahydrofolate production, leading to DNA damage, stress response induction, and accumulation of reactive oxygen species (ROS) in a process known as thymineless death (TLD). TLD survival can occur through the uptake of exogenous thymidine, countering the effects of SXT; however, a growing body of research has implicated central metabolism as another potentially important determinant of bacterial survival of SXT and other antibiotics. Here, we conducted studies to better understand the mechanisms of TLD survival in S. aureus. We found that thymidine abundances in CF sputum were insufficient to prevent TLD of S. aureus, highlighting the importance of alternative survival mechanisms in vivo. In S. aureus cultured in vitro with SXT and low thymidine, we frequently identified adaptive mutations in genes encoding carbohydrate, nucleotide, and amino acid metabolism, supporting reduced metabolism as a common survival mechanism. Although intracellular ROS levels rose with SXT treatment in vitro, survival was not improved in the presence of ROS scavengers, unlike in E. coli. SXT challenge induced the SOS response, which was alleviated by added thymidine. Finally, an inactivating mutation in the phosphotransferase gene ptsI conferred both limitation in cellular ATP and improved survival against TLD. Collectively, these results suggest that alterations in core metabolic functions, particularly those that reduce ATP levels, predominantly confer S. aureus survival and persistence during SXT treatment, potentially identifying novel targets for co-treatment.IMPORTANCEStaphylococcus aureus is a ubiquitous organism and one of the leading causes of human infections, many of which are difficult to treat due to persistence, antibiotic resistance, or antibiotic tolerance. As our arsenal of effective antibiotics dwindles, the need for improved treatments becomes increasingly urgent, necessitating a better understanding of the precise mechanisms by which pathogens evade our most critical antimicrobial agents. Here, we report a systematic characterization of the mechanisms of S. aureus survival to treatment with the first-line antistaphylococcal antibiotic trimethoprim-sulfamethoxazole, identifying pathways and candidate targets for enhancing the efficacy of available antimicrobial agents.https://journals.asm.org/doi/10.1128/mbio.01634-24Staphylococcus aureusthymineless deathantibiotic resistanceantifolate drugspersistence
spellingShingle Lauren J. Gonsalves
Allyson Tran
Tessa Gardiner
Tiia Freeman
Angshita Dutta
Carson J. Miller
Sharon McNamara
Adam Waalkes
Dustin R. Long
Stephen J. Salipante
Lucas R. Hoffman
Daniel J. Wolter
Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
mBio
Staphylococcus aureus
thymineless death
antibiotic resistance
antifolate drugs
persistence
title Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
title_full Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
title_fullStr Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
title_full_unstemmed Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
title_short Mechanisms of Staphylococcus aureus survival of trimethoprim-sulfamethoxazole-induced thymineless death
title_sort mechanisms of staphylococcus aureus survival of trimethoprim sulfamethoxazole induced thymineless death
topic Staphylococcus aureus
thymineless death
antibiotic resistance
antifolate drugs
persistence
url https://journals.asm.org/doi/10.1128/mbio.01634-24
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