The ΔCysK mutant of Mycobacterium tuberculosis is sensitive to vancomycin associated with changes in cell wall phospholipid profile

Emani, Carine Sao; Richter, Adrian; Singh, Ajit; Bhatt, Apoorva; Av‐Gay, Yossef

doi:10.1016/j.bbrc.2022.07.096

Cited by 7 publications

(7 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Assortment of DEGs into functional classes revealed intermediary metabolism & respiration and cell wall pathways among the most impacted categories (Figure 2h). These results corroborate the recent finding that CysK2 alters the phospholipid profile of the Mtb cell envelope [11]. Pathway enrichment analysis of the most enriched Gene ontology biological process indicated that while most pathways are commonly affected by CysM and CysK2, these L-cysteine synthases also have a unique transcriptional footprint (Table S4).…”

Section: Resultssupporting

confidence: 88%

“…Unlike Mtb Δ cysH , which was reported to be an L-cysteine auxotroph and thus required L-methionine or glutathione supplementation to grow in vitro (from which L-cysteine can be generated catabolically)[15], neither deletion of cysK2 nor cysM impacted Mtb growth kinetics in vitro suggesting that these genes are functionally redundant during rich growth conditions [5] [11] (Figure S4). Interestingly, the induction of various host-like stresses attenuated the growth of these mutants compared to the parental strain, pointing towards the possibility of an enhanced requirement of L-cysteine-derived antioxidants and other biomolecules to subvert these stresses (Figure 1).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Divergent downstream biosynthetic pathways are supported by L-cysteine synthases ofMycobacterium tuberculosis

Khan,

Hunt,

Singha

et al. 2023

Preprint

View full text Add to dashboard Cite

Mycobacterium tuberculosis’s (Mtb)autarkic lifestyle within the host involves rewiring its transcriptional networks to combat host-induced stresses. With the help of RNA-seq performed under various stress conditions, we identified that genes belonging toMtbsulfur metabolism pathways are significantly upregulated during oxidative stress. Using an integrated approach of microbial genetics, transcriptomics, metabolomics, animal experiments, chemical inhibition, and rescue studies, we investigated the biological role of non-canonical L-cysteine synthases, CysM and CysK2. While transcriptome signatures ofRvΔcysMandRvΔcysK2appear similar under regular growth conditions, we observed unique transcriptional signatures when subjected to oxidative stress. We followed pool size and labelling (34S) of key downstream metabolites, viz. mycothiol and ergothioneine, to monitor L-cysteine biosynthesis and utilization. This revealed the significant role of distinct L-cysteine biosynthetic routes on redox stress and homeostasis. CysM and CysK2 independently facilitateMtbsurvival by alleviating host-induced redox stress, suggesting they are not fully redundant during infection. With the help of genetic mutants and chemical inhibitors, we show that CysM and CysK2 serve as unique, attractive targets for adjunct therapy to combat mycobacterial infection.

show abstract

Section: Resultssupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Divergent downstream biosynthetic pathways are supported by L-cysteine synthases ofMycobacterium tuberculosis

Khan,

Hunt,

Singha

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…It has a growth defect in Sauton’s media, in THP1 macrophages and is sensitive to SDS and copper ( Sakthi and Narayanan, 2013 ). Similarly, the ∆ rv0848 (∆ cysK 2 ) mutant was shown to have an altered cell wall lipid profile, to be sensitive to oxidative stress, vancomycin, rifampicin and have a growth defect in Raw264.7 macrophages ( Sao Emani et al, 2022 ). The gene rv0849 codes for an MFS-type efflux pump.…”

Section: Resultsmentioning

confidence: 99%

“…By using a range of concentrations of CuOOH, we investigated the expression profile of rv0191 along with that of rv1877 , rv1878 , and rv0848 ( cysK 2 ). The gene rv0848 was previously shown to be upregulated during oxidative stress ( Voskuil et al, 2011 ) and to be required for the protection of M.tb against OS ( Sao Emani et al, 2022 ). As opposed to the expression of cysK 2 that remained high in all tested concentrations of CuOOH, the expression of rv0191 , rv1877 and rv1878 was not altered ( Supplementary Figure S4 ), indicating that the ROS detoxification role of Rv0191 is distinct to that of Rv0848.…”

Section: Resultsmentioning

confidence: 99%

“…Growth curves were obtained by sub-culturing a starter cultures (initiated from frozen stocks) in 7H9-ADS-Tyl [supplemented with 1X albumin-dextrose-sodium chloride (ADS) and 0.06% tyloxapol as previously described ( Sao Emani et al, 2018d , 2022 )]. The preparation of the 10X ADS supplement consisted to add 25 g Bovine Albumin Fraction V (Sigma Aldrich), 10 g Dextrose, (Sigma Aldrich) and 4.25 g Sodium Chloride (Sigma Aldrich) to a final volume of 490 mL distilled water.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The efflux pumps Rv1877 and Rv0191 play differential roles in the protection of Mycobacterium tuberculosis against chemical stress

Sao Emani,

Reiling

2024

Front. Microbiol.

Self Cite

View full text Add to dashboard Cite

BackgroundIt was previously shown that GlnA3sc enabled Streptomyces coelicolor to survive in excess polyamines. However, subsequent studies revealed that Rv1878, the corresponding Mycobacterium tuberculosis (M.tb) ortholog, was not essential for the detoxification of spermine (Spm), in M.tb. On the other hand, the multi-drug efflux pump Rv1877 was previously shown to enable export of a wide range of compounds, while Rv0191 was shown to be more specific to chloramphenicol.RationaleTherefore, we first wanted to determine if detoxification of Spm by efflux can be achieved by any efflux pump, or if that was dependent upon the function of the pump. Next, since Rv1878 was found not to be essential for the detoxification of Spm, we sought to follow-up on the investigation of the physiological role of Rv1878 along with Rv1877 and Rv0191.ApproachTo evaluate the specificity of efflux pumps in the mycobacterial tolerance to Spm, we generated unmarked ∆rv1877 and ∆rv0191 M.tb mutants and evaluated their susceptibility to Spm. To follow up on the investigation of any other physiological roles they may have, we characterized them along with the ∆rv1878 M.tb mutant.ResultsThe ∆rv1877 mutant was sensitive to Spm stress, while the ∆rv0191 mutant was not. On the other hand, the ∆rv1878 mutant grew better than the wild-type during iron starvation yet was sensitive to cell wall stress. The proteins Rv1877 and Rv1878 seemed to play physiological roles during hypoxia and acidic stress. Lastly, the ∆rv0191 mutant was the only mutant that was sensitive to oxidative stress.ConclusionThe multidrug MFS-type efflux pump Rv1877 is required for Spm detoxification, as opposed to Rv0191 which seems to play a more specific role. Moreover, Rv1878 seems to play a role in the regulation of iron homeostasis and the reconstitution of the cell wall of M.tb. On the other hand, the sensitivity of the ∆rv0191 mutant to oxidative stress, suggests that Rv0191 may be responsible for the transport of low molecular weight thiols.

show abstract

Mycobacterium tuberculosis and its clever approaches to escape the deadly macrophage

Krishnan,

Nath,

Nair

et al. 2023

World J Microbiol Biotechnol

View full text Add to dashboard Cite

The ΔCysK mutant of Mycobacterium tuberculosis is sensitive to vancomycin associated with changes in cell wall phospholipid profile

Cited by 7 publications

References 41 publications

Divergent downstream biosynthetic pathways are supported by L-cysteine synthases ofMycobacterium tuberculosis

Divergent downstream biosynthetic pathways are supported by L-cysteine synthases ofMycobacterium tuberculosis

The efflux pumps Rv1877 and Rv0191 play differential roles in the protection of Mycobacterium tuberculosis against chemical stress

Mycobacterium tuberculosis and its clever approaches to escape the deadly macrophage

Contact Info

Product

Resources

About