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Many plant bacterial pathogens including  Pseudomonas  species, utilize the type III secretion system (T3SS) to deliver effector proteins into plant cells. Genes encoding the T3SS and its effectors are repressed in nutrient-rich media but are rapidly induced after the bacteria enter a plant or are transferred into nutrient-deficient media. To understand how the T3SS genes are regulated, we screened for  P .  savastanoi  pv.  phaseolicola  ( Psph ) mutants displaying diminished induction of  avrPto-luc , a reporter for the T3SS genes, in  Arabidopsis . A mutant carrying transposon insertion into a gene coding for a small functional unknown protein, designated as  rhpC , was identified that poorly induced  avrPto-luc  in plants and in minimal medium (MM). Interestingly,  rhpC  is located immediately downstream of a putative metalloprotease gene named  rhpP , and the two genes are organized in an operon  rhpPC ; but  rhpP  and  rhpC  displayed different RNA expression patterns in nutrient-rich King’s B medium (KB) and MM. Deletion of the whole  rhpPC  locus did not significantly affect the  avrPto-luc  induction, implying coordinated actions of  rhpP  and  rhpC  in regulating the T3SS genes. Further analysis showed that RhpC was a cytoplasmic protein that interacted with RhpP and targeted RhpP to the periplasm. In the absence of RhpC, RhpP was localized in the cytoplasm and caused a reduction of HrpL, a key regulator of the T3SS genes, and also reduced the fitness of  Psph . Expression of RhpP alone in  E .  coli  inhibited the bacterial growth. The detrimental effect of RhpP on the fitness of  Psph  and  E .  coli  required metalloprotease active sites, and was repressed when RhpC was co-expressed with RhpP. The coordination between  rhpP  and  rhpC  in tuning the T3SS gene expression and cell fitness reveals a novel regulatory mechanism for bacterial pathogenesis. The function of RhpP in the periplasm remains to be studied.

plos pathogens

Two components of the rhpPC operon coordinately regulate the type III secretion system and bacterial fitness in Pseudomonas savastanoi pv. phaseolicola