Characterisation of a putative M23-domain containing protein in Mycobacterium tuberculosis.
Andrea Olga PapadopoulosChristopher EalandBhavna Gowan GordhanMichael VanNieuwenhzeBavesh Davandra KanaPublished in: PloS one (2021)
Mycobacterium tuberculosis, the causative agent of tuberculosis remains a global health concern, further compounded by the high rates of HIV-TB co-infection and emergence of multi- and extensive drug resistant TB, all of which have hampered efforts to eradicate this disease. As a result, novel anti-tubercular interventions are urgently required, with the peptidoglycan component of the M. tuberculosis cell wall emerging as an attractive drug target. Peptidoglycan M23 endopeptidases can function as active cell wall hydrolases or degenerate activators of hydrolases in a variety of bacteria, contributing to important processes such as bacterial growth, division and virulence. Herein, we investigate the function of the Rv0950-encoded putative M23 endopeptidase in M. tuberculosis. In silico analysis revealed that this protein is conserved in mycobacteria, with a zinc-binding catalytic site predictive of hydrolytic activity. Transcript analysis indicated that expression of Rv0950c was elevated during lag and log phases of growth and reduced in stationary phase. Deletion of Rv0950c yielded no defects in growth, colony morphology, antibiotic susceptibility or intracellular survival but caused a reduction in cell length. Staining with a monopeptide-derived fluorescent D-amino acid, which spatially reports on sites of active PG biosynthesis or repair, revealed an overall reduction in uptake of the probe in ΔRv0950c. When stained with a dipeptide probe in the presence of cell wall damaging agents, the ΔRv0950c mutant displayed reduced sidewall labelling. As bacterial peptidoglycan metabolism is important for survival and pathogenesis, the role of Rv0950c and other putative M23 endopeptidases in M. tuberculosis should be explored further.
Keyphrases
- mycobacterium tuberculosis
- cell wall
- drug resistant
- pulmonary tuberculosis
- amino acid
- global health
- single cell
- multidrug resistant
- quantum dots
- binding protein
- living cells
- poor prognosis
- escherichia coli
- acinetobacter baumannii
- pseudomonas aeruginosa
- human immunodeficiency virus
- public health
- hepatitis c virus
- antiretroviral therapy
- hiv infected
- free survival
- protein protein
- transcription factor
- emergency department
- physical activity
- stem cells
- hiv testing
- staphylococcus aureus
- rna seq
- men who have sex with men
- long non coding rna
- cell therapy
- mass spectrometry
- small molecule
- bacillus subtilis
- fluorescent probe
- wild type
- label free
- single molecule