Identification of Novel Efflux Proteins Rv0191, Rv3756c, Rv3008, and Rv1667c Involved in Pyrazinamide Resistance in Mycobacterium tuberculosis.

Abstract

Pyrazinamide (PZA) is a critical drug used for the treatment of tuberculosis (TB). PZA is a prodrug that requires conversion to the active component pyrazinoic acid (POA) by pyrazinamidase (PZase) encoded by the pncA gene. Although resistance to PZA is mostly caused by pncA mutations and less commonly by rpsA, panD, and clpC1 mutations, clinical strains without these mutations are known to exist. While efflux of POA was demonstrated in Mycobacterium tuberculosis previously, the efflux proteins involved have not been identified. Here we performed POA binding studies with an M. tuberculosis proteome microarray and identified four efflux proteins (Rv0191, Rv3756c, Rv3008, and Rv1667c) that bind POA. Overexpression of the four efflux pump genes in M. tuberculosis caused low-level resistance to PZA and POA but not to other drugs. Furthermore, addition of efflux pump inhibitors such as reserpine, piperine, and verapamil caused increased susceptibility to PZA in M. tuberculosis strains overexpressing the efflux proteins Rv0191, Rv3756c, Rv3008, and Rv1667c. Our studies indicate that these four efflux proteins may be responsible for PZA/POA efflux and cause PZA resistance in M. tuberculosis Future studies are needed to assess their roles in PZA resistance in clinical strains.

• Authors: Yumeng Zhang, Jia Zhang, Peng Cui, Ying Zhang, Wenhong Zhang
• Journal: Antimicrobial agents and chemotherapy (Antimicrob Agents Chemother) Vol. 61 Issue 8 (08 2017) ISSN: 1098-6596 [Electronic] United States
• PMID: 28584158 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2017 American Society for Microbiology.

Chemical References

• Alkaloids
• Antitubercular Agents
• Benzodioxoles
• Carrier Proteins
• Membrane Transport Proteins
• Piperidines
• Polyunsaturated Alkamides
• Pyrazinamide
• pyrazinoic acid
• Reserpine
• Verapamil
• Amidohydrolases
• PncA protein, Mycobacterium tuberculosis
• piperine

Topics

• Alkaloids (pharmacology)
• Amidohydrolases (genetics)
• Antitubercular Agents (pharmacology)
• Benzodioxoles (pharmacology)
• Carrier Proteins (antagonists & inhibitors)
• Drug Resistance, Bacterial (genetics)
• Humans
• Membrane Transport Proteins (biosynthesis, genetics)
• Mycobacterium tuberculosis (drug effects, genetics)
• Piperidines (pharmacology)
• Polyunsaturated Alkamides (pharmacology)
• Pyrazinamide (analogs & derivatives, metabolism, pharmacology)
• Reserpine (pharmacology)
• Tuberculosis, Pulmonary (drug therapy, microbiology)
• Verapamil (pharmacology)