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Metallochaperones Are Needed for Mycobacterium tuberculosis and Escherichia coli Nicotinamidase-Pyrazinamidase Activity.

Sheen, P; Monsalve, A; Campos, J; Huerta, R; Antiparra, R; Arteaga, H; Duran, P; Bueno, C; Kirwan, DE; Gilman, RH; et al. Sheen, P; Monsalve, A; Campos, J; Huerta, R; Antiparra, R; Arteaga, H; Duran, P; Bueno, C; Kirwan, DE; Gilman, RH; Zimic, M (2020) Metallochaperones Are Needed for Mycobacterium tuberculosis and Escherichia coli Nicotinamidase-Pyrazinamidase Activity. J Bacteriol, 202 (2). ISSN 1098-5530 https://doi.org/10.1128/JB.00331-19
SGUL Authors: Kirwan, Daniela Elisa

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Abstract

Mycobacterium tuberculosis nicotinamidase-pyrazinamidase (PZAse) is a metalloenzyme that catalyzes conversion of nicotinamide-pyrazinamide to nicotinic acid-pyrazinoic acid. This study investigated whether a metallochaperone is required for optimal PZAse activity. M. tuberculosis and Escherichia coli PZAses (PZAse-MT and PZAse-EC, respectively) were inactivated by metal depletion (giving PZAse-MT-Apo and PZAse-EC-Apo). Reactivation with the E. coli metallochaperone ZnuA or Rv2059 (the M. tuberculosis analog) was measured. This was repeated following proteolytic and thermal treatment of ZnuA and Rv2059. The CDC1551 M. tuberculosis reference strain had the Rv2059 coding gene knocked out, and PZA susceptibility and the pyrazinoic acid (POA) efflux rate were measured. ZnuA (200 μM) achieved 65% PZAse-EC-Apo reactivation. Rv2059 (1 μM) and ZnuA (1 μM) achieved 69% and 34.3% PZAse-MT-Apo reactivation, respectively. Proteolytic treatment of ZnuA and Rv2059 and application of three (but not one) thermal shocks to ZnuA significantly reduced the capacity to reactivate PZAse-MT-Apo. An M. tuberculosis Rv2059 knockout strain was Wayne positive and susceptible to PZA and did not have a significantly different POA efflux rate than the reference strain, although a trend toward a lower efflux rate was observed after knockout. The metallochaperone Rv2059 restored the activity of metal-depleted PZAse in vitro Although Rv2059 is important in vitro, it seems to have a smaller effect on PZA susceptibility in vivo. It may be important to mechanisms of action and resistance to pyrazinamide in M. tuberculosis Further studies are needed for confirmation.IMPORTANCE Tuberculosis is an infectious disease caused by the bacterium Mycobacterium tuberculosis and remains one of the major causes of disease and death worldwide. Pyrazinamide is a key drug used in the treatment of tuberculosis, yet its mechanism of action is not fully understood, and testing strains of M. tuberculosis for pyrazinamide resistance is not easy with the tools that are presently available. The significance of the present research is that a metallochaperone-like protein may be crucial to pyrazinamide's mechanisms of action and of resistance. This may support the development of improved tools to detect pyrazinamide resistance, which would have significant implications for the clinical management of patients with tuberculosis: drug regimens that are appropriately tailored to the resistance profile of a patient's individual strain lead to better clinical outcomes, reduced onward transmission of infection, and reduction of the development of resistant strains that are more challenging and expensive to treat.

Item Type: Article
Additional Information: Copyright © 2020 Sheen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/).
Keywords: Mycobacterium tuberculosis, PZA resistance, Rv2059, ZnuA, mechanism of action, mechanism of resistance, metal depletion, metallochaperone, metalloenzyme, pyrazinamide, pyrazinoic acid, reactivation, resistance, Mycobacterium tuberculosis, PZA resistance, pyrazinamide, pyrazinoic acid, metallochaperone, metalloenzyme, resistance, mechanism of action, mechanism of resistance, ZnuA, Rv2059, reactivation, metal depletion, 06 Biological Sciences, 11 Medical and Health Sciences, 07 Agricultural and Veterinary Sciences, Microbiology
SGUL Research Institute / Research Centre: Academic Structure > Infection and Immunity Research Institute (INII)
Journal or Publication Title: J Bacteriol
ISSN: 1098-5530
Language: eng
Dates:
DateEvent
2 January 2020Published
4 October 2019Accepted
Publisher License: Creative Commons: Attribution 4.0
Projects:
Project IDFunderFunder ID
099805/Z/12/ZWellcome Trusthttp://dx.doi.org/10.13039/100004440
K23 NR018482NINR NIH HHSUNSPECIFIED
0687-01-10Grand Challenge CanadaUNSPECIFIED
037-2014FONDECyT-PeruUNSPECIFIED
PubMed ID: 31636108
Web of Science ID: WOS:000505200900001
Go to PubMed abstract
URI: http://openaccess.sgul.ac.uk/id/eprint/111568
Publisher's version: https://doi.org/10.1128/JB.00331-19

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