Analysis of uracil phosphoribosyltransferase expression in Mycobacterium tuberculosis and evaluation of upp knockout strain in infected mice

FEMS Microbiol Lett. 2017 Feb 1;364(4). doi: 10.1093/femsle/fnx023.

Abstract

The upp (Rv3309c)-encoded uracil phosphoribosyltransferase from Mycobacterium tuberculosis (MtUPRT) converts uracil and 5-phosphoribosyl-α-1-pyrophosphate into pyrophosphate and uridine 5΄-monophosphate, the precursor of all pyrimidine nucleotides. A M. tuberculosis knockout strain for upp gene was generated by allelic replacement. Knockout and complemented strains were validated by a functional assay of uracil incorporation. A basal level of MtUPRT expression is shown to be independent of either growth medium used, addition of bases, or oxygen presence/absence. The upp disruption does not affect M. tuberculosis growth in Middlebrook 7H9 medium, and it is not required for M. tuberculosis virulence in a mouse model of infection. Thus, MtUPRT is unlikely to be a good target for drugs against M. tuberculosis.

Keywords: Mycobacterium tuberculosis; Wayne model; gene knockout; mouse model; upp gene; uracil phosphoribosyltransferase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Disease Models, Animal
  • Gene Expression*
  • Gene Knockout Techniques
  • Mice
  • Mycobacterium tuberculosis / enzymology*
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / growth & development
  • Mycobacterium tuberculosis / pathogenicity*
  • Pentosyltransferases / genetics*
  • Pentosyltransferases / metabolism
  • Tuberculosis / microbiology*
  • Uracil / metabolism
  • Uracil / pharmacology
  • Virulence

Substances

  • Bacterial Proteins
  • Uracil
  • Pentosyltransferases
  • uracil phosphoribosyltransferase