The sensor kinase MprB is required for Rhodococcus equi virulence.
Rhodococcus equi is a soil bacterium and, like Mycobacterium tuberculosis, a member of the mycolata. Through possession of a virulence plasmid, it has the ability to infect the alveolar macrophages of foals, resulting in pyogranulomatous bronchopneumonia. The virulence plasmid has an orphan two-component system (TCS) regulatory gene, orf8, mutation of which completely attenuates virulence. This study attempted to find the cognate sensor kinase (SK) of orf8. Annotation of the R. equi strain 103 genome identified 23 TCSs encoded on the chromosome, which were used in a DNA microarray to compare TCS gene transcription in murine macrophage-like cells to growth in vitro. This identified six SKs as significantly up-regulated during growth in macrophages. Mutants of these SKs were constructed and their ability to persist in macrophages was determined with one SK, MprB, found to be required for intracellular survival. The attenuation of the mprB- mutant, and its complementation, was confirmed in a mouse virulence assay. In silico analysis of the R. equi genome sequence identified an MprA binding box motif homologous to that of M. tuberculosis, on mprA, pepD, sigB and sigE. The results of this study also show that R. equi responds to the macrophage environment differently from M. tuberculosis. MprB is the first SK identified as required for R. equi virulence and intracellular survival.
MacArthur
Iain
I
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Gordon Street, Guelph, Ontario, N1G 2W1, Canada.
Parreira
Valeria R
VR
Lepp
Dion
D
Mutharia
Lucy M
LM
Vazquez-Boland
José A
JA
Prescott
John F
JF
D000193:Actinomycetales Infections / Q000382:microbiology* / Q000401:mortality; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000662:veterinary; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D016929:Rhodococcus equi / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D016415:Sequence Alignment; D014774:Virulence / Q000235:genetics*
D011494:Protein Kinases