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A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish.

Mohanty, Soumitra; Jagannathan, Lakshmanan; Ganguli, Geetanjali; Padhi, Avinash; Roy, Debasish; Alaridah, Nader LU ; Saha, Pratip; Nongthomba, Upendra; Godaly, Gabriela LU and Kumar, Gopal Ramesh, et al. (2015) In Journal of Biological Chemistry 290(21). p.13321-13343
Abstract
Mycobacterium tuberculosis (Mtb) employs various strategies to modulate host immune responses to facilitate its persistence in macrophages. The Mtb cell wall contains numerous glycoproteins with unknown role in pathogenesis. Here, by using concavalinA and LC-MS analysis we identified a novel mannosylated glycoprotein phosphoribosyl- transferase, encoded by the Rv3242c, from Mtb cell walls. Homology modeling, bioinformatic analyses and assay of phosphoribosyltransferase activity measurement in Mycobacterium smegmatis expressing recombinant Rv3242c (MsmRv3242c) confirmed the mass spectrometry data. Using Mycobacterium marinum-zebrafish and the surrogate MsmRv3242c infection models, we proved that phosphoribosyltransferase is involved in... (More)
Mycobacterium tuberculosis (Mtb) employs various strategies to modulate host immune responses to facilitate its persistence in macrophages. The Mtb cell wall contains numerous glycoproteins with unknown role in pathogenesis. Here, by using concavalinA and LC-MS analysis we identified a novel mannosylated glycoprotein phosphoribosyl- transferase, encoded by the Rv3242c, from Mtb cell walls. Homology modeling, bioinformatic analyses and assay of phosphoribosyltransferase activity measurement in Mycobacterium smegmatis expressing recombinant Rv3242c (MsmRv3242c) confirmed the mass spectrometry data. Using Mycobacterium marinum-zebrafish and the surrogate MsmRv3242c infection models, we proved that phosphoribosyltransferase is involved in mycobacterial virulence. Histological and infection assays showed that M. marinum mimG mutant, an Rv3242c orthologue in a pathogenic M. marinum strain, was strongly attenuated in adult zebrafish and also survived less in macrophages. In contrast, infection with wild-type and the complemented ∆mimG:Rv3242c M. marinum strains showed prominent pathological features such as severe emaciation, skin lesions, hemorrhaging, and more zebrafish death. Similarly, recombinant MsmRv3242c bacteria showed increased invasion in non-phagocytic epithelial cells and longer intracellular survival in macrophages as compared to wild-type and vector control M. smegmatis strains. Further mechanistic studies revealed that the Rv3242c and mimG mediated enhancement of intramacrophagic survival was due to inhibition of autophagy, reactive oxygen species and reduced activities of superoxide dismutase and catalase enzymes. Infection with MsmRv3242c also activated the MAPK pathway, NF-κB and inflammatory cytokines. In summary, we show that a novel mycobacterial mannosylated phosphoribosyltransferase acts as a virulence and immunomodulatory factors, suggesting that it may constitute a novel target for antimycobacterial drugs. (Less)
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Journal of Biological Chemistry
volume
290
issue
21
pages
13321 - 13343
publisher
ASBMB
external identifiers
  • pmid:25825498
  • wos:000354975700031
  • scopus:84929997383
ISSN
1083-351X
DOI
10.1074/jbc.M114.598482
language
English
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ff41753a-bcb4-475f-8e08-d08984f30c0a (old id 5360714)
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http://www.ncbi.nlm.nih.gov/pubmed/25825498?dopt=Abstract
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2015-05-07 19:23:47
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@article{ff41753a-bcb4-475f-8e08-d08984f30c0a,
  abstract     = {Mycobacterium tuberculosis (Mtb) employs various strategies to modulate host immune responses to facilitate its persistence in macrophages. The Mtb cell wall contains numerous glycoproteins with unknown role in pathogenesis. Here, by using concavalinA and LC-MS analysis we identified a novel mannosylated glycoprotein phosphoribosyl- transferase, encoded by the Rv3242c, from Mtb cell walls. Homology modeling, bioinformatic analyses and assay of phosphoribosyltransferase activity measurement in Mycobacterium smegmatis expressing recombinant Rv3242c (MsmRv3242c) confirmed the mass spectrometry data. Using Mycobacterium marinum-zebrafish and the surrogate MsmRv3242c infection models, we proved that phosphoribosyltransferase is involved in mycobacterial virulence. Histological and infection assays showed that M. marinum mimG mutant, an Rv3242c orthologue in a pathogenic M. marinum strain, was strongly attenuated in adult zebrafish and also survived less in macrophages. In contrast, infection with wild-type and the complemented ∆mimG:Rv3242c M. marinum strains showed prominent pathological features such as severe emaciation, skin lesions, hemorrhaging, and more zebrafish death. Similarly, recombinant MsmRv3242c bacteria showed increased invasion in non-phagocytic epithelial cells and longer intracellular survival in macrophages as compared to wild-type and vector control M. smegmatis strains. Further mechanistic studies revealed that the Rv3242c and mimG mediated enhancement of intramacrophagic survival was due to inhibition of autophagy, reactive oxygen species and reduced activities of superoxide dismutase and catalase enzymes. Infection with MsmRv3242c also activated the MAPK pathway, NF-κB and inflammatory cytokines. In summary, we show that a novel mycobacterial mannosylated phosphoribosyltransferase acts as a virulence and immunomodulatory factors, suggesting that it may constitute a novel target for antimycobacterial drugs.},
  author       = {Mohanty, Soumitra and Jagannathan, Lakshmanan and Ganguli, Geetanjali and Padhi, Avinash and Roy, Debasish and Alaridah, Nader and Saha, Pratip and Nongthomba, Upendra and Godaly, Gabriela and Kumar, Gopal Ramesh and Banerjee, Sulagna and Sonawane, Avinash},
  issn         = {1083-351X},
  language     = {eng},
  number       = {21},
  pages        = {13321--13343},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish.},
  url          = {http://dx.doi.org/10.1074/jbc.M114.598482},
  volume       = {290},
  year         = {2015},
}