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Published on: March 28, 2025
Characterization of a chromosomal toxin-antitoxin, Rv1102c-Rv1103c system in Mycobacterium tuberculosis
Jeong-Sun Han1, Jae Jin Lee, Tripti Anandan
1Department of Biological Sciences, Myongji University, San 38-2 Namdong, Yongin, Gyeonggido 449-728, Republic of Korea.
Abstract:
Toxin-antitoxin systems, ubiquitous in prokaryotic genomes, have been proposed to play an important role in several stress responses. While Mycobacterium tuberculosis contains more than 80 putative TA loci, the roles they play in this pathogen are yet to be studied. Here, we characterize a chromosomal Rv1102c-Rv1103c TA system in M. tuberculosis. We found that the Rv1102c toxin interacts with the Rv1103c antitoxin in a pull-down assay and the yeast two-hybrid system. Rv1102c cleaved the era mRNA in Escherichia coli, and cleavage was inhibited by co-expression of Rv1103c. Heterologous expression of Rv1102c led to growth arrest in E. coli, which was fully recovered only when Rv1103c was co-expressed in cis with Rv1102c, suggesting that the production and assembly of Rv1102c and Rv1103c are tightly linked. Our additional results indicate that translational coupling of the Rv1102c and Rv1103c genes is important for Rv1102c-Rv1103c binding. Finally, we discovered that the expression of Rv1102c induced growth arrest and increased the level of persister cells in Mycobacterium smegmatis. These results suggest that the Rv1102c-Rv1103c TA system could play a role in M. tuberculosis pathogenesis via generating bacilli that survive in the face of multidrug therapy.
Insights
The Rv1102c-Rv1103c toxin-antitoxin system in Mycobacterium tuberculosis is characterized. This system
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Toxin-antitoxin (TA) systems are prevalent in prokaryotes and implicated in stress responses.
- Mycobacterium tuberculosis possesses over 80 putative TA loci, but their functions remain largely unelucidated.
- Understanding TA systems is crucial for deciphering bacterial survival mechanisms.
Purpose of the Study:
- To characterize the Rv1102c-Rv1103c chromosomal toxin-antitoxin system in Mycobacterium tuberculosis.
- To investigate the functional interaction between Rv1102c (toxin) and Rv1103c (antitoxin).
- To explore the role of this TA system in bacterial growth, survival, and pathogenesis.
Main Methods:
- Protein-protein interaction assays (pull-down, yeast two-hybrid).
- mRNA cleavage assays in Escherichia coli.
- Heterologous gene expression and growth analysis in E. coli and Mycobacterium smegmatis.
- Analysis of translational coupling between Rv1102c and Rv1103c genes.
Main Results:
- Rv1102c toxin directly interacts with Rv1103c antitoxin.
- Rv1102c cleaves era mRNA, an activity inhibited by Rv1103c.
- Heterologous expression of Rv1102c causes growth arrest in E. coli, rescued by co-expressed Rv1103c.
- Translational coupling of Rv1102c-Rv1103c genes is vital for their interaction.
- Rv1102c expression induces growth arrest and increases persister cell levels in Mycobacterium smegmatis.
Conclusions:
- The Rv1102c-Rv1103c TA system is functionally characterized in M. tuberculosis.
- This TA system influences bacterial growth and survival under stress.
- The Rv1102c-Rv1103c system may contribute to M. tuberculosis pathogenesis by promoting survival during multidrug therapy.
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