Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase
Abstract Background Curiosity on toxin–antitoxin modules has increased intensely over recent years as it is ubiquitously present in many bacterial genomes, including pathogens like Methicillin-resistant Staphylococcus aureus (MRSA). Several cellular functions of TA systems have been proposed however...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-05-01
|
| Series: | Journal of Biomedical Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12929-022-00810-5 |
| _version_ | 1818200849257594880 |
|---|---|
| author | Sonia Jain Arghya Bhowmick Bohyun Jeong Taeok Bae Abhrajyoti Ghosh |
| author_facet | Sonia Jain Arghya Bhowmick Bohyun Jeong Taeok Bae Abhrajyoti Ghosh |
| author_sort | Sonia Jain |
| collection | DOAJ |
| description | Abstract Background Curiosity on toxin–antitoxin modules has increased intensely over recent years as it is ubiquitously present in many bacterial genomes, including pathogens like Methicillin-resistant Staphylococcus aureus (MRSA). Several cellular functions of TA systems have been proposed however, their exact role in cellular physiology remains unresolved. Methods This study aims to find out the impact of the mazEF toxin–antitoxin module on biofilm formation, pathogenesis, and antibiotic resistance in an isolated clinical ST239 MRSA strain, by constructing mazE and mazF mutants using CRISPR–cas9 base-editing plasmid (pnCasSA-BEC). Transcriptome analysis (RNA-seq) was performed for the mazE antitoxin mutant in order to identify the differentially regulated genes. The biofilm formation was also assessed for the mutant strains. Antibiogram profiling was carried out for both the generated mutants followed by murine experiment to determine the pathogenicity of the constructed strains. Results For the first time our work showed, that MazF promotes cidA mediated cell death and lysis for biofilm formation without playing any significant role in host virulence as suggested by the murine experiment. Interestingly, the susceptibility to oxacillin, daptomycin and vancomycin was reduced significantly by the activated MazF toxin in the mazE mutant strain. Conclusions Our study reveals that activated MazF toxin leads to resistance to antibiotics like oxacillin, daptomycin and vancomycin. Therefore, in the future, any potential antibacterial drug can be designed to target MazF toxin against the problematic multi-drug resistant bug. |
| first_indexed | 2024-12-12T02:44:11Z |
| format | Article |
| id | doaj.art-50a5167581f44d6a8e94ae4c0669922b |
| institution | Directory Open Access Journal |
| issn | 1423-0127 |
| language | English |
| last_indexed | 2024-12-12T02:44:11Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biomedical Science |
| spelling | doaj.art-50a5167581f44d6a8e94ae4c0669922b2022-12-22T00:41:05ZengBMCJournal of Biomedical Science1423-01272022-05-0129111610.1186/s12929-022-00810-5Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminaseSonia Jain0Arghya Bhowmick1Bohyun Jeong2Taeok Bae3Abhrajyoti Ghosh4Infectious Disease and Immunology Division, CSIR-Indian Institute of Chemical BiologyDepartment of Biochemistry, Bose InstituteDepartment of Microbiology, Kosin University College of MedicineDepartment of Microbiology and Immunology, Indiana University, School of Medicine-NorthwestDepartment of Biochemistry, Bose InstituteAbstract Background Curiosity on toxin–antitoxin modules has increased intensely over recent years as it is ubiquitously present in many bacterial genomes, including pathogens like Methicillin-resistant Staphylococcus aureus (MRSA). Several cellular functions of TA systems have been proposed however, their exact role in cellular physiology remains unresolved. Methods This study aims to find out the impact of the mazEF toxin–antitoxin module on biofilm formation, pathogenesis, and antibiotic resistance in an isolated clinical ST239 MRSA strain, by constructing mazE and mazF mutants using CRISPR–cas9 base-editing plasmid (pnCasSA-BEC). Transcriptome analysis (RNA-seq) was performed for the mazE antitoxin mutant in order to identify the differentially regulated genes. The biofilm formation was also assessed for the mutant strains. Antibiogram profiling was carried out for both the generated mutants followed by murine experiment to determine the pathogenicity of the constructed strains. Results For the first time our work showed, that MazF promotes cidA mediated cell death and lysis for biofilm formation without playing any significant role in host virulence as suggested by the murine experiment. Interestingly, the susceptibility to oxacillin, daptomycin and vancomycin was reduced significantly by the activated MazF toxin in the mazE mutant strain. Conclusions Our study reveals that activated MazF toxin leads to resistance to antibiotics like oxacillin, daptomycin and vancomycin. Therefore, in the future, any potential antibacterial drug can be designed to target MazF toxin against the problematic multi-drug resistant bug.https://doi.org/10.1186/s12929-022-00810-5MRSABiofilmAntibiotic resistanceVirulenceCRISPR–Cas9Toxin–antitoxin |
| spellingShingle | Sonia Jain Arghya Bhowmick Bohyun Jeong Taeok Bae Abhrajyoti Ghosh Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase Journal of Biomedical Science MRSA Biofilm Antibiotic resistance Virulence CRISPR–Cas9 Toxin–antitoxin |
| title | Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase |
| title_full | Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase |
| title_fullStr | Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase |
| title_full_unstemmed | Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase |
| title_short | Unravelling the physiological roles of mazEF toxin–antitoxin system on clinical MRSA strain by CRISPR RNA-guided cytidine deaminase |
| title_sort | unravelling the physiological roles of mazef toxin antitoxin system on clinical mrsa strain by crispr rna guided cytidine deaminase |
| topic | MRSA Biofilm Antibiotic resistance Virulence CRISPR–Cas9 Toxin–antitoxin |
| url | https://doi.org/10.1186/s12929-022-00810-5 |
| work_keys_str_mv | AT soniajain unravellingthephysiologicalrolesofmazeftoxinantitoxinsystemonclinicalmrsastrainbycrisprrnaguidedcytidinedeaminase AT arghyabhowmick unravellingthephysiologicalrolesofmazeftoxinantitoxinsystemonclinicalmrsastrainbycrisprrnaguidedcytidinedeaminase AT bohyunjeong unravellingthephysiologicalrolesofmazeftoxinantitoxinsystemonclinicalmrsastrainbycrisprrnaguidedcytidinedeaminase AT taeokbae unravellingthephysiologicalrolesofmazeftoxinantitoxinsystemonclinicalmrsastrainbycrisprrnaguidedcytidinedeaminase AT abhrajyotighosh unravellingthephysiologicalrolesofmazeftoxinantitoxinsystemonclinicalmrsastrainbycrisprrnaguidedcytidinedeaminase |