VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis
The prokaryotic ubiquitous Toxin-antitoxin (TA) modules encodes for a stable toxin and an unstable antitoxin. VapBC subfamily is the most abundant Type II TA system in M. tuberculosis genome. However, the exact physiological role for most of these Type II TA systems are still unknown. Here, we have...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2020.02037/full |
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| author | Arun Sharma Gopinath Chattopadhyay Pankaj Chopra Munmun Bhasin Chandrani Thakur Sakshi Agarwal Shahbaz Ahmed Nagasuma Chandra Raghavan Varadarajan Raghavan Varadarajan Ramandeep Singh |
| author_facet | Arun Sharma Gopinath Chattopadhyay Pankaj Chopra Munmun Bhasin Chandrani Thakur Sakshi Agarwal Shahbaz Ahmed Nagasuma Chandra Raghavan Varadarajan Raghavan Varadarajan Ramandeep Singh |
| author_sort | Arun Sharma |
| collection | DOAJ |
| description | The prokaryotic ubiquitous Toxin-antitoxin (TA) modules encodes for a stable toxin and an unstable antitoxin. VapBC subfamily is the most abundant Type II TA system in M. tuberculosis genome. However, the exact physiological role for most of these Type II TA systems are still unknown. Here, we have comprehensively characterized the VapBC21 TA locus from M. tuberculosis. The overexpression of VapC21 inhibited mycobacterial growth in a bacteriostatic manner and as expected, growth inhibition was abrogated upon co-expression of the cognate antitoxin, VapB21. We observed that the deletion of vapC21 had no noticeable influence on the in vitro and in vivo growth of M. tuberculosis. Using co-expression and biophysical studies, we observed that in addition to VapB21, VapC21 is also able to interact with non-cognate antitoxin, VapB32. The strength of interaction varied between the cognate and non-cognate TA pairs. The overexpression of VapC21 resulted in differential expression of approximately 435 transcripts in M. tuberculosis. The transcriptional profiles obtained upon ectopic expression of VapC21 was similar to those reported in M. tuberculosis upon exposure to stress conditions such as nutrient starvation and enduring hypoxic response. Further, VapC21 overexpression also led to increased expression of WhiB7 regulon and bacterial tolerance to aminoglycosides and ethambutol. Taken together, these results indicate that a complex network of interactions exists between non-cognate TA pairs and VapC21 contributes to drug tolerance in vitro. |
| first_indexed | 2024-12-20T21:38:12Z |
| format | Article |
| id | doaj.art-6ce0bffdfe1545b882b277f460bd530e |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-20T21:38:12Z |
| publishDate | 2020-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-6ce0bffdfe1545b882b277f460bd530e2022-12-21T19:25:53ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2020-09-011110.3389/fmicb.2020.02037559879VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosisArun Sharma0Gopinath Chattopadhyay1Pankaj Chopra2Munmun Bhasin3Chandrani Thakur4Sakshi Agarwal5Shahbaz Ahmed6Nagasuma Chandra7Raghavan Varadarajan8Raghavan Varadarajan9Ramandeep Singh10Tuberculosis Research Laboratory, Translational Health Science and Technology Institute, Faridabad, IndiaMolecular Biophysics Unit, Indian Institute of Science, Bengaluru, IndiaTuberculosis Research Laboratory, Translational Health Science and Technology Institute, Faridabad, IndiaMolecular Biophysics Unit, Indian Institute of Science, Bengaluru, IndiaDepartment of Biochemistry, Indian Institute of Science, Bengaluru, IndiaTuberculosis Research Laboratory, Translational Health Science and Technology Institute, Faridabad, IndiaMolecular Biophysics Unit, Indian Institute of Science, Bengaluru, IndiaDepartment of Biochemistry, Indian Institute of Science, Bengaluru, IndiaMolecular Biophysics Unit, Indian Institute of Science, Bengaluru, IndiaJawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, IndiaTuberculosis Research Laboratory, Translational Health Science and Technology Institute, Faridabad, IndiaThe prokaryotic ubiquitous Toxin-antitoxin (TA) modules encodes for a stable toxin and an unstable antitoxin. VapBC subfamily is the most abundant Type II TA system in M. tuberculosis genome. However, the exact physiological role for most of these Type II TA systems are still unknown. Here, we have comprehensively characterized the VapBC21 TA locus from M. tuberculosis. The overexpression of VapC21 inhibited mycobacterial growth in a bacteriostatic manner and as expected, growth inhibition was abrogated upon co-expression of the cognate antitoxin, VapB21. We observed that the deletion of vapC21 had no noticeable influence on the in vitro and in vivo growth of M. tuberculosis. Using co-expression and biophysical studies, we observed that in addition to VapB21, VapC21 is also able to interact with non-cognate antitoxin, VapB32. The strength of interaction varied between the cognate and non-cognate TA pairs. The overexpression of VapC21 resulted in differential expression of approximately 435 transcripts in M. tuberculosis. The transcriptional profiles obtained upon ectopic expression of VapC21 was similar to those reported in M. tuberculosis upon exposure to stress conditions such as nutrient starvation and enduring hypoxic response. Further, VapC21 overexpression also led to increased expression of WhiB7 regulon and bacterial tolerance to aminoglycosides and ethambutol. Taken together, these results indicate that a complex network of interactions exists between non-cognate TA pairs and VapC21 contributes to drug tolerance in vitro.https://www.frontiersin.org/article/10.3389/fmicb.2020.02037/fullMycobacterium tuberculosistoxin-antitoxin systemribonucleasedrug tolerancecross talk |
| spellingShingle | Arun Sharma Gopinath Chattopadhyay Pankaj Chopra Munmun Bhasin Chandrani Thakur Sakshi Agarwal Shahbaz Ahmed Nagasuma Chandra Raghavan Varadarajan Raghavan Varadarajan Ramandeep Singh VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis Frontiers in Microbiology Mycobacterium tuberculosis toxin-antitoxin system ribonuclease drug tolerance cross talk |
| title | VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis |
| title_full | VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis |
| title_fullStr | VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis |
| title_full_unstemmed | VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis |
| title_short | VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis |
| title_sort | vapc21 toxin contributes to drug tolerance and interacts with non cognate vapb32 antitoxin in mycobacterium tuberculosis |
| topic | Mycobacterium tuberculosis toxin-antitoxin system ribonuclease drug tolerance cross talk |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2020.02037/full |
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