The pncA gene from naturally pyrazinamide-resistant Mycobacterium avium encodes pyrazinamidase and confers pyrazinamide susceptibility to resistant M. tuberculosis complex organisms

Microbiology (Reading). 1997 Oct;143 ( Pt 10):3367-73. doi: 10.1099/00221287-143-10-3367.

Abstract

The antituberculosis drug pyrazinamide (PZA) needs to be converted into pyrazinoic acid (POA) by the bacterial pyrazinamidase (PZase) in order to show bactericidal activity against Mycobacterium tuberculosis. M. avium is naturally resistant to PZA. To investigate whether this natural resistance to PZA is due to inability of the M. avium PZase to convert PZA to bactericidal POA, the M. avium PZase gene (pncA) was cloned by using the M. tuberculosis pncA gene as a probe. Sequence analysis showed that the M. avium pncA gene is 561 bp long, encoding a protein with a predicted size of about 19.8 kDa; but Western blotting showed that the M. avium PZase migrated as a 24 kDa band when expressed in M. bovis BCG and Escherichia coli. Sequence comparison revealed that M. avium PZase has 67.7% and 32.8% amino acid identity with the corresponding enzymes from M. tuberculosis and E. coli, respectively. Southern blot analysis with the M. avium pncA gene as a probe showed that M. terrae, M. gastri, M. marinum, M. fortuitum, M. xenopi, M. gordonae, M. szulgai, M. celatum and M. kansasii have close pncA homologues, whereas M. chelonae and M. smegmatis did not give significant hybridization signals. Transformation with the M. avium pncA gene conferred PZA susceptibility to PZA-resistant M. tuberculosis complex organisms, indicating that the nonsusceptibility of M. avium to PZA is not due to an ineffective PZase enzyme, but appears to be related to other factors such as transport of POA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / genetics*
  • Amidohydrolases / metabolism
  • Amino Acid Sequence
  • Antitubercular Agents / metabolism
  • Antitubercular Agents / pharmacology
  • Base Sequence
  • Biological Transport, Active / genetics
  • Cloning, Molecular
  • DNA Primers / genetics
  • Drug Resistance, Microbial / genetics
  • Escherichia coli / genetics
  • Genes, Bacterial*
  • Humans
  • Molecular Sequence Data
  • Mycobacterium / enzymology
  • Mycobacterium / genetics
  • Mycobacterium avium Complex / drug effects
  • Mycobacterium avium Complex / enzymology*
  • Mycobacterium avium Complex / genetics*
  • Pyrazinamide / analogs & derivatives
  • Pyrazinamide / metabolism
  • Pyrazinamide / pharmacology
  • Sequence Homology, Amino Acid
  • Species Specificity

Substances

  • Antitubercular Agents
  • DNA Primers
  • Pyrazinamide
  • pyrazinoic acid
  • Amidohydrolases
  • pyrazinamide deamidase

Associated data

  • GENBANK/U80820