Depletion of M. tuberculosis GlmU from Infected Murine Lungs Effects the Clearance of the Pathogen.
M. tuberculosis N-acetyl-glucosamine-1-phosphate uridyltransferase (GlmUMtb) is a bi-functional enzyme engaged in the synthesis of two metabolic intermediates N-acetylglucosamine-1-phosphate (GlcNAc-1-P) and UDP-GlcNAc, catalyzed by the C- and N-terminal domains respectively. UDP-GlcNAc is a key metabolite essential for the synthesis of peptidoglycan, disaccharide linker, arabinogalactan and mycothiols. While glmUMtb was predicted to be an essential gene, till date the role of GlmUMtb in modulating the in vitro growth of Mtb or its role in survival of pathogen ex vivo / in vivo have not been deciphered. Here we present the results of a comprehensive study dissecting the role of GlmUMtb in arbitrating the survival of the pathogen both in vitro and in vivo. We find that absence of GlmUMtb leads to extensive perturbation of bacterial morphology and substantial reduction in cell wall thickness under normoxic as well as hypoxic conditions. Complementation studies show that the acetyl- and uridyl- transferase activities of GlmUMtb are independently essential for bacterial survival in vitro, and GlmUMtb is also found to be essential for mycobacterial survival in THP-1 cells as well as in guinea pigs. Depletion of GlmUMtb from infected murine lungs, four weeks post infection, led to significant reduction in the bacillary load. The administration of Oxa33, a novel oxazolidine derivative that specifically inhibits GlmUMtb, to infected mice resulted in significant decrease in the bacillary load. Thus our study establishes GlmUMtb as a strong candidate for intervention measures against established tuberculosis infections.
National Institute of Immunology, New Delhi, India; Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad Campus, Hyderabad, India.
Vijay
V
Soni
National Institute of Immunology, New Delhi, India.
Sandeep
S
Upadhayay
Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad Campus, Hyderabad, India.
Priyanka
P
Suryadevara
Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad Campus, Hyderabad, India.
Ganesh
G
Samla
CSIR-Institute of Genomics and Integrative Biology, New Delhi, India.
Archana
A
Singh
Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad Campus, Hyderabad, India.
Perumal
P
Yogeeswari
Department of Pharmacy, Birla Institute of Technology and Science-Pilani, Hyderabad Campus, Hyderabad, India.
Dharmarajan
D
Sriram
National Institute of Immunology, New Delhi, India.
Vinay Kumar
VK
Nandicoori
D000818:Animals; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors*; D001597:Benzylidene Compounds / Q000494:pharmacology; D015153:Blotting, Western; D004195:Disease Models, Animal; D006168:Guinea Pigs; D051379:Mice; D008807:Mice, Inbred BALB C; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D016297:Mutagenesis, Site-Directed; D009169:Mycobacterium tuberculosis; D009281:Naphthalenes / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D014376:Tuberculosis / Q000378:metabolism*
D001426:Bacterial Proteins; D001597:Benzylidene Compounds; C532100:GlmU protein, Mycobacterium tuberculosis; D009097:Multienzyme Complexes; D009281:Naphthalenes; D010080:Oxazoles