The vapBC Operon from Mycobacterium smegmatis Is An Autoregulated Toxin–Antitoxin Module That Controls Growth via Inhibition of Translation

Robson, Jennifer R.; McKenzie, Joanna Leigh; Cursons, Raymond T.; Cook, Gregory M.; Arcus, Vickery L.

doi:10.1016/j.jmb.2009.05.006

Cited by 96 publications

(95 citation statements)

References 46 publications

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“…Strain RF100 was transformed with pJR113 (22), and the mutant was created as described previously, and this is strain RF102. This strain was then transformed with pRF221, and the triple mutant (⌬TA triple ; RF105) was created using the two-step crossover protocol.…”

Section: Methodsmentioning

confidence: 99%

“…The orientation of the insert was checked using primers phdlac F and 3ЈmcspUC R. This plasmid was then electroporated into M. smegmatis mc 2 155 and strain RF101, creating strains RF121 and RF122, respectively. ␤-Galactosidase assays were carried out as described previously (22).…”

Section: Methodsmentioning

confidence: 99%

“…MSMEG_1282 was up-regulated in ⌬vapBC 3-fold, but the reason for this is unknown. MSMEG_1282 has been shown to be part of a separate mRNA transcript to the vapBC TA system (22). However, the vapBC operon is autoregulated, and the promoter region overlaps with MSMEG_1282, so in the deletion strain the removal of the VapBC complex from binding to that region of the DNA may allow some up-regulation.…”

Section: Smegmatis Vapbc-mazef-phd/doc Triple Deletion Mutant Is Dmentioning

confidence: 99%

“…The number of TA systems in the genome of Mycobacterium tuberculosis has been greatly expanded with 88 putative TA systems present (19,21). In contrast, the chromosome of Mycobacterium leprae contains only toxin pseudogenes, whereas Mycobacterium smegmatis contains only three TA systems (19,22). The greatest number of TA modules in M. tuberculosis belongs to VapBC with 47 in total.…”

mentioning

confidence: 99%

“…Extracellular samples were additionally normalized by the relative abundance of metabolite identified in the uncultured medium. Finally, the data were log-transformed to fit the (22) and two other putative TA operons, mazEF and phd/doc (19). MazF from M. smegmatis (MSMEG_4448) is a member of the PemK superfamily (NCBI annotation ϭ transcriptional modulator of MazE; Pfam number ϭ PF02452), which currently contains 1393 proteins from 376 species of bacteria and archaea.…”

mentioning

confidence: 99%

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Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Frampton

Aggio

Villas‐Bôas

et al. 2012

Journal of Biological Chemistry

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Background: Mycobacteria harbor a vast array of toxin-antitoxin modules, but their roles remain largely unknown. Results: Deletion of all TA modules in Mycobacterium smegmatis caused a survival defect and alterations in amino acid metabolism. Conclusion:We demonstrate an essential role for TA modules in mycobacterial metabolism and survival. Significance: These results may explain the basis for 88 TA modules in M. tuberculosis where metabolism must be tightly controlled.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Smegmatis Vapbc-mazef-phd/doc Triple Deletion Mutant Is Dmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Frampton

Aggio

Villas‐Bôas

et al. 2012

Journal of Biological Chemistry

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Transcriptional Control of Toxin–Antitoxin Expression: Keeping Toxins Under Wraps Until the Time is Right

Kędzierska

Hayes

2016

Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria

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The crystal structure of the Rv0301‐Rv0300 VapBC‐3 toxin—antitoxin complex from M. tuberculosis reveals a Mg²⁺ ion in the active site and a putative RNA‐binding site

et al. 2012

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VapBC pairs account for 45 out of 88 identified toxin-antitoxin (TA) pairs in the Mycobacterium tuberculosis (Mtb) H37Rv genome. A working model suggests that under times of stress, antitoxin molecules are degraded, releasing the toxins to slow the metabolism of the cell, which in the case of VapC toxins is via their RNase activity. Otherwise the TA pairs remain bound to their promoters, autoinhibiting transcription. The crystal structure of Rv0301-Rv0300, an Mtb VapBC TA complex determined at 1.49 Å resolution, suggests a mechanism for these three functions: RNase activity, its inhibition by antitoxin, and its ability to bind promoter DNA. The Rv0301 toxin consists of a core of five parallel beta strands flanked by alpha helices. Three proximal aspartates coordinate a Mg 21 ion forming the putative RNase active site. The Rv0300 antitoxin monomer is extended in structure, consisting of an N-terminal beta strand followed by four helices. The last two helices wrap around the toxin and terminate near the putative RNase active site, but with different conformations. In one conformation, the C-terminal arginine interferes with Mg 21 ion coordination, suggesting a mechanism by which the antitoxin can inhibit toxin activity. At the N-terminus of the antitoxin, two pairs of Ribbon-Helix-Helix (RHH) motifs are related by crystallographic twofold symmetry. The resulting hetero-octameric complex is similar to the FitAB system, but the two RHH motifs are about 30 Å closer together in the Rv0301-Rv0300 complex, suggesting either a different span of the DNA recognition sequence or a conformational change.

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The vapBC Operon from Mycobacterium smegmatis Is An Autoregulated Toxin–Antitoxin Module That Controls Growth via Inhibition of Translation

Cited by 96 publications

References 46 publications

Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Transcriptional Control of Toxin–Antitoxin Expression: Keeping Toxins Under Wraps Until the Time is Right

The crystal structure of the Rv0301‐Rv0300 VapBC‐3 toxin—antitoxin complex from M. tuberculosis reveals a Mg²⁺ ion in the active site and a putative RNA‐binding site

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The vapBC Operon from Mycobacterium smegmatis Is An Autoregulated Toxin–Antitoxin Module That Controls Growth via Inhibition of Translation

Cited by 96 publications

References 46 publications

Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Toxin-Antitoxin Systems of Mycobacterium smegmatis Are Essential for Cell Survival

Transcriptional Control of Toxin–Antitoxin Expression: Keeping Toxins Under Wraps Until the Time is Right

The crystal structure of the Rv0301‐Rv0300 VapBC‐3 toxin—antitoxin complex from M. tuberculosis reveals a Mg2+ ion in the active site and a putative RNA‐binding site

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The crystal structure of the Rv0301‐Rv0300 VapBC‐3 toxin—antitoxin complex from M. tuberculosis reveals a Mg²⁺ ion in the active site and a putative RNA‐binding site