RfiA, a novel PAP2 domain-containing polytopic membrane protein that confers resistance to the FtsZ inhibitor PC190723

Future Microbiol. 2015;10(3):325-35. doi: 10.2217/fmb.14.131.

Abstract

Background: As an essential protein for bacterial cell division, the tubulin-like FtsZ protein has been selected as a target for development of next generation antimicrobials. PC190723 is a fluoride-containing benzamide compound developed as a FtsZ inhibitor that selectively inhibits growth of multidrug resistant Gram-positive bacteria.

Aim: Our aim was to investigate the mechanism of resistance to PC109723 conferred by over-expression of a gene, rfiA, in an environmental bacterium Arthrobacter A3.

Materials & methods: The investigations included analysis of the effect of PC109723 on wild-type Arthrobacter A3 and a recombinant strain over-expressing rfiA, in vivo localization of RfiA, in vitro measurements of fluorine release from PC109723 by membrane extracts from the over-expression strain combined with mass spectrophotometric analysis of reaction products, and modelling of RfiA structure.

Results: We describe a novel protein, RfiA, from Arthrobacter A3 that confers PC190723 resistance. RfiA is a PAP2 domain-containing polytopic transmembrane protein that can modify the fluoridated benzamide ring that is critical for high affinity binding of PC190723 with FtsZ.

Conclusion: RfiA-mediated modification of PC190723 is the first reported instance of resistance to this antibiotic involving a change to its structure. We predict that adoption of PC190723 or related benzamides as antimicrobials in clinical practice will lead to the acquisition by resistant pathogens of a gene encoding this subfamily of proteins.

Keywords: FtsZ; antibiotic resistance; cell cytoskeleton; efflux; mechanisms of resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arthrobacter / drug effects*
  • Arthrobacter / genetics
  • Arthrobacter / metabolism
  • Bacterial Proteins / antagonists & inhibitors
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism*
  • Benzamides / metabolism
  • Cytoskeletal Proteins / antagonists & inhibitors*
  • Cytoskeletal Proteins / metabolism
  • Drug Resistance, Bacterial / genetics*
  • Membrane Proteins / genetics
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism*
  • Microbial Sensitivity Tests
  • Phylogeny
  • Pyridines / pharmacology*
  • Thiazoles / pharmacology*

Substances

  • Bacterial Proteins
  • Benzamides
  • Cytoskeletal Proteins
  • FtsZ protein, Bacteria
  • Membrane Proteins
  • PC190723
  • Pyridines
  • Thiazoles
  • benzamide