PUBLICATION
A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish
- Authors
- Mohanty, S., Jagannathan, L., Ganguli, G., Padhi, A., Roy, D., Alaridah, N., Saha, P., Nongthomba, U., Godaly, G., Kumar, G.R., Banerjee, S., Sonawane, A.
- ID
- ZDB-PUB-150401-5
- Date
- 2015
- Source
- The Journal of biological chemistry 290(21): 13321-43 (Journal)
- Registered Authors
- Keywords
- Mycobacterium marinum, Mycobacterium smegmatis, Mycobacterium tuberculosis, autophagy, macrophage, mimG, mitogen-activated protein kinase (MAPK), oxidative stress, phosphoribosyltransferase, zebrafish
- MeSH Terms
-
- Animals
- Apoptosis
- Autophagy*
- Blotting, Western
- Cell Adhesion
- Cell Movement
- Cell Proliferation
- Cell Wall/metabolism
- Cells, Cultured
- Female
- Host-Pathogen Interactions
- Humans
- Macrophages/cytology
- Macrophages/immunology*
- Macrophages/microbiology
- Mice
- Mice, Inbred BALB C
- Microbial Viability
- Mycobacterium marinum/growth & development
- Mycobacterium marinum/pathogenicity*
- Mycobacterium tuberculosis/growth & development
- Mycobacterium tuberculosis/pathogenicity*
- NF-kappa B
- Nicotinamide Phosphoribosyltransferase/genetics
- Nicotinamide Phosphoribosyltransferase/metabolism*
- Oxidative Stress*
- Phagocytosis
- Protein Conformation
- RNA, Messenger/genetics
- Reactive Oxygen Species/metabolism
- Real-Time Polymerase Chain Reaction
- Reverse Transcriptase Polymerase Chain Reaction
- Spectrometry, Mass, Electrospray Ionization
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Superoxide Dismutase/genetics
- Superoxide Dismutase/metabolism
- Tuberculosis/immunology*
- Tuberculosis/metabolism
- Tuberculosis/microbiology
- Virulence/immunology
- Zebrafish/immunology*
- Zebrafish/metabolism
- Zebrafish/microbiology
- PubMed
- 25825498 Full text @ J. Biol. Chem.
Citation
Mohanty, S., Jagannathan, L., Ganguli, G., Padhi, A., Roy, D., Alaridah, N., Saha, P., Nongthomba, U., Godaly, G., Kumar, G.R., Banerjee, S., Sonawane, A. (2015) A mycobacterial phosphoribosyltransferase promotes bacillary survival by inhibiting oxidative stress and autophagy pathways in macrophages and zebrafish. The Journal of biological chemistry. 290(21):13321-43.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping