Inorganic polyphosphate accumulation suppresses the dormancy response and virulence in Mycobacterium tuberculosis

Tiwari, Priya; Gosain, Tannu Priya; Singh, Mamta; Sankhe, Gaurav D.; Arora, Garima; Kidwai, Saqib; Agarwal, Sakshi; Chugh, Saurabh; Saini, Deepak Kumar; Singh, Ramandeep

doi:10.1074/jbc.ra119.008370

Cited by 13 publications

(12 citation statements)

References 70 publications

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“…We hypothesize that this may be explained by the fact of slow growing mycobacteria requiring DosR to metabolically adapt to low O 2 levels as already reported 27 . This result is also in agreement with reduced dosR transcription and reduced biofilm production observed in a double mutant devoid of the exopolyphosphatases genes 28 and also in a mtrB mutant 29 .…”

Section: Discussionsupporting

confidence: 89%

Transcriptional portrait of M. bovis BCG during biofilm production shows genes differentially expressed during intercellular aggregation and substrate attachment

Flores-Valdéz

Aceves-Sánchez

Peterson

et al. 2020

Sci Rep

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Mycobacterium tuberculosis and M. smegmatis form drug-tolerant biofilms through dedicated genetic programs. In support of a stepwise process regulating biofilm production in mycobacteria, it was shown elsewhere that lsr2 participates in intercellular aggregation, while groEL1 was required for biofilm maturation in M. smegmatis. Here, by means of RNA-Seq, we monitored the early steps of biofilm production in M. bovis BCG, to distinguish intercellular aggregation from attachment to a surface. Genes encoding for the transcriptional regulators dosR and BCG0114 (Rv0081) were significantly regulated and responded differently to intercellular aggregation and surface attachment. Moreover, a M. tuberculosis H37Rv deletion mutant in the Rv3134c-dosS-dosR regulon, formed less biofilm than wild type M. tuberculosis, a phenotype reverted upon reintroduction of this operon into the mutant. Combining RT-qPCR with microbiological assays (colony and surface pellicle morphologies, biofilm quantification, Ziehl-Neelsen staining, growth curve and replication of planktonic cells), we found that BCG0642c affected biofilm production and replication of planktonic BcG, whereas ethR affected only phenotypes linked to planktonic cells despite its downregulation at the intercellular aggregation step. our results provide evidence for a stage-dependent expression of genes that contribute to biofilm production in slow-growing mycobacteria. In nature, microbial species are often found within a matrix, forming multicellular communities that attach to surfaces or air-liquid interfaces, called biofilms 1. Biofilms are relevant to human health, as a majority of bacterial pathogens employ these structures to modify the host response 2 contributing to persistence 3. In this regard, a link exists between in vitro biofilm production and in vivo persistence for BCG 4 and M. tuberculosis 5. Biofilm formation occurs via a series of well-defined steps. These include the attachment of single-cell planktonic microbes onto a substratum; aggregation and growth of the adherent cells into three-dimensionally organized structures; and encapsulation of the structures by a self-produced matrix of extracellular polymeric substance 1 .

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Section: Discussionsupporting

confidence: 89%

Transcriptional portrait of M. bovis BCG during biofilm production shows genes differentially expressed during intercellular aggregation and substrate attachment

Flores-Valdéz

Aceves-Sánchez

Peterson

et al. 2020

Sci Rep

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“…The colony morphology and ability of these strains to form biofilms was determined as previously described (Arora et al, 2018;Tiwari et al, 2019). The effect of deletion of vapC21 on the survival of M. tuberculosis was evaluated in the following conditions: oxidative stress (5 mM H 2 O 2 for 3 days), nitrosative stress (5 mM NaNO 2 , pH-5.2 for 3 days), 0.25% SDS for 3 days, 2.5 mg/ml lysozyme for 3 days, nutrient starvation (1× Tris buffered saline, 1× TBS with 0.05% Tween-80 for 7 and 14 days), drugs (isoniazid, rifampicin, and levofloxacin for 14 days) as previously described (Arora et al, 2018;Tiwari et al, 2019). For bacterial enumeration, at designated time points, 10-fold serial dilutions were prepared and plated on Middlebrook 7H11 medium 37 • C for 3-4 weeks.…”

Section: In Vitro Stress and In Vivo Experimentsmentioning

confidence: 99%

“…The isolated RNA was subjected to DNase I treatment, cDNA libraries were prepared from rRNA depleted samples and sequenced using a Illumina HiSeq platform at Aggrigenome labs Pvt. Ltd. as previously described (Tiwari et al, 2019). The data obtained from the mutant strain was analyzed as previously mentioned (Tiwari et al, 2019).…”

Section: Rna Sequencing Experimentsmentioning

confidence: 99%

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VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis

et al. 2020

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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.

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“…54,55 In M tuberculosis, polyphosphate plays roles in infection persistence, survival, and virulence, while in T cruzi, polyphosphate is important for environmental stress adaptation. 54,[56][57][58] Tuberculosis infection can lead to the development of pulmonary fibrosis, whereas T cruzi infection can lead to the development of cardiac fibrosis. [59][60][61] Microbial polyphosphate may therefore be a source of this fibrosis-inducing activity, which warrants further investigation.…”

Section: Discussionmentioning

confidence: 99%

Platelet polyphosphate induces fibroblast chemotaxis and myofibroblast differentiation

Suess

Smith

Morrissey

2020

Journal of Thrombosis and Haemostasis

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Background: Platelets secrete many pro-wound healing molecules such as growth factors and cytokines. We found that releasates from activated human platelets induced the differentiation of cultured murine and human fibroblasts into a myofibroblast phenotype. Surprisingly, most of this differentiation-inducing activity was heat-stable, suggesting it was not due to the protein component of the releasates. Inorganic polyphosphate is a major constituent of platelet-dense granules and promotes blood coagulation and inflammation. Objectives: We aim to investigate the contribution of polyphosphate on myofibroblast differentiating activity of platelet releasates. Methods: Using NIH-3T3 cells and primary human fibroblasts, we examined the effect of human platelet releasates and chemically synthesized polyphosphate on fibroblast differentiation and migration. Results: We found that the myofibroblast-inducing activity of platelet releasates was severely attenuated after incubation with a polyphosphate-degrading enzyme, and that fibroblasts responded to platelet-sized polyphosphate by increased levels of α-smooth muscle actin, stress fibers, and collagen. Furthermore, fibroblasts were chemotactic toward polyphosphate. Conclusions: These findings indicate that platelet-derived polyphosphate acts as a cell signaling molecule by inducing murine and human fibroblasts to differentiate into myofibroblasts, a cell type known to drive both wound healing and fibrosing diseases. Polyphosphate therefore not only promotes early wound responses through enhancing fibrin clot formation, but also may play roles in the later stages of wound healing, and, potentially, progression of fibrotic diseases, by recruiting fibroblasts and inducing their differentiation into myofibroblasts.

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Inorganic polyphosphate accumulation suppresses the dormancy response and virulence in Mycobacterium tuberculosis

Cited by 13 publications

References 70 publications

Transcriptional portrait of M. bovis BCG during biofilm production shows genes differentially expressed during intercellular aggregation and substrate attachment

Transcriptional portrait of M. bovis BCG during biofilm production shows genes differentially expressed during intercellular aggregation and substrate attachment

VapC21 Toxin Contributes to Drug-Tolerance and Interacts With Non-cognate VapB32 Antitoxin in Mycobacterium tuberculosis

Platelet polyphosphate induces fibroblast chemotaxis and myofibroblast differentiation

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