Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis

Pyrazinamide (PZA) is a frontline anti-tuberculosis drug that plays a crucial role in the treatment of both drug-susceptible and multidrug-resistant tuberculosis (MDR-TB). PZA is a prodrug that is converted to its active form, pyrazinoic acid (POA), by a nicotinamidase/pyrazinamidase encoded by the...

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Main Authors:	Shi, Wanliang, Chen, Jiazhen, Feng, Jie, Cui, Peng, Zhang, Shuo, Weng, Xinhua, Zhang, Wenhong, Zhang, Ying
Format:	Online
Language:	English
Published:	Nature Publishing Group 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4150287/

id	pubmed-4150287
recordtype	oai_dc
spelling	pubmed-41502872014-09-03 Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis Shi, Wanliang Chen, Jiazhen Feng, Jie Cui, Peng Zhang, Shuo Weng, Xinhua Zhang, Wenhong Zhang, Ying Original Article Pyrazinamide (PZA) is a frontline anti-tuberculosis drug that plays a crucial role in the treatment of both drug-susceptible and multidrug-resistant tuberculosis (MDR-TB). PZA is a prodrug that is converted to its active form, pyrazinoic acid (POA), by a nicotinamidase/pyrazinamidase encoded by the pncA gene, the mutation of which is the major cause of PZA resistance. Although RpsA (ribosomal protein S1, involved in trans-translation) has recently been shown to be a target of POA/PZA, whole-genome sequencing has identified mutations in the panD gene encoding aspartate decarboxylase in PZA-resistant strains lacking pncA and rpsA mutations. To gain more insight into a possible new target of PZA, we isolated 30 POA-resistant mutants lacking mutations in pncA and rpsA from M. tuberculosis in vitro, and whole-genome sequencing of 3 mutants identified various mutations in the panD gene. Additionally, sequencing analysis revealed that the remaining 27 POA-resistant mutants all harbored panD mutations affecting the C-terminus of the PanD protein, with PanD M117I being the most frequent mutation (24/30, 80%). Conditional overexpression of panD from M. tuberculosis, M. smegmatis or E. coli, or of M. tuberculosis mutant PanD M117I, all conferred resistance to POA and PZA in M. tuberculosis. β-alanine and pantothenate, which are downstream products of PanD, were found to antagonize the antituberculosis activity of POA. In addition, the activity of the M. tuberculosis PanD enzyme was inhibited by POA at therapeutically relevant concentrations in a concentration-dependent manner but was not inhibited by the prodrug PZA or the control compound nicotinamide. These findings suggest that PanD represents a new target of PZA/POA. These results have implications for a better understanding of this peculiar persister drug and for the design of new drugs targeting M. tuberculosis persisters for improved treatment. Nature Publishing Group 2014-08 2014-08-13 /pmc/articles/PMC4150287/ /pubmed/26038753 http://dx.doi.org/10.1038/emi.2014.61 Text en Copyright © 2014 Shanghai Shangyixun Cultural Communication Co., Ltd http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Shi, Wanliang Chen, Jiazhen Feng, Jie Cui, Peng Zhang, Shuo Weng, Xinhua Zhang, Wenhong Zhang, Ying
spellingShingle	Shi, Wanliang Chen, Jiazhen Feng, Jie Cui, Peng Zhang, Shuo Weng, Xinhua Zhang, Wenhong Zhang, Ying Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
author_facet	Shi, Wanliang Chen, Jiazhen Feng, Jie Cui, Peng Zhang, Shuo Weng, Xinhua Zhang, Wenhong Zhang, Ying
author_sort	Shi, Wanliang
title	Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
title_short	Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
title_full	Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
title_fullStr	Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
title_full_unstemmed	Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis
title_sort	aspartate decarboxylase (pand) as a new target of pyrazinamide in mycobacterium tuberculosis
description	Pyrazinamide (PZA) is a frontline anti-tuberculosis drug that plays a crucial role in the treatment of both drug-susceptible and multidrug-resistant tuberculosis (MDR-TB). PZA is a prodrug that is converted to its active form, pyrazinoic acid (POA), by a nicotinamidase/pyrazinamidase encoded by the pncA gene, the mutation of which is the major cause of PZA resistance. Although RpsA (ribosomal protein S1, involved in trans-translation) has recently been shown to be a target of POA/PZA, whole-genome sequencing has identified mutations in the panD gene encoding aspartate decarboxylase in PZA-resistant strains lacking pncA and rpsA mutations. To gain more insight into a possible new target of PZA, we isolated 30 POA-resistant mutants lacking mutations in pncA and rpsA from M. tuberculosis in vitro, and whole-genome sequencing of 3 mutants identified various mutations in the panD gene. Additionally, sequencing analysis revealed that the remaining 27 POA-resistant mutants all harbored panD mutations affecting the C-terminus of the PanD protein, with PanD M117I being the most frequent mutation (24/30, 80%). Conditional overexpression of panD from M. tuberculosis, M. smegmatis or E. coli, or of M. tuberculosis mutant PanD M117I, all conferred resistance to POA and PZA in M. tuberculosis. β-alanine and pantothenate, which are downstream products of PanD, were found to antagonize the antituberculosis activity of POA. In addition, the activity of the M. tuberculosis PanD enzyme was inhibited by POA at therapeutically relevant concentrations in a concentration-dependent manner but was not inhibited by the prodrug PZA or the control compound nicotinamide. These findings suggest that PanD represents a new target of PZA/POA. These results have implications for a better understanding of this peculiar persister drug and for the design of new drugs targeting M. tuberculosis persisters for improved treatment.
publisher	Nature Publishing Group
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4150287/
_version_	1613129670833733632

Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis

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