Regulation of Iron Transport inStreptococcus pneumoniaeby RitR, an Orphan Response Regulator

Open Access
Publisher Website
Google Scholar
Abstract
RitR (formerly RR489) is an orphan two-component signal transduction response regulator inStreptococcus pneumoniaethat has been shown to be required for lung pathogenicity. In the present study, by using the rough strain R800, inactivation of the orphan response regulator generitRby allele replacement reduced pathogenicity in a cyclophosphamide-treated mouse lung model but not in a thigh model, suggesting a role for RitR in regulation of tissue-specific virulence factors. Analysis of changes in genome-wide transcript mRNA levels associated with the inactivation ofritRcompared to wild-type cells was performed by the use of high-density DNA microarrays. Genes with a change in transcript abundance associated with inactivation ofritRincludedpiuB, encoding an Fe permease subunit, andpiuA, encoding an Fe carrier-binding protein. In addition, adprortholog, encoding an H2O2resistance protein that has been shown to reduce synthesis of reactive oxygen intermediates, was activated in the wild-type (ritR+) strain. Microarray experiments suggested that RitR represses Fe uptake in vitro by negatively regulating the Piu hemin-iron transport system. Footprinting experiments confirmed site-specific DNA-binding activity for RitR and identified three binding sites that partly overlap the +1 site for transcription initiation upstream ofpiuB. Transcripts belonging to other gene categories found to be differentially expressed in our array studies include those associated with (i) H2O2resistance, (ii) repair of DNA damage, (iii) sugar transport and capsule biosynthesis, and (iv) two-component signal transduction elements. These observations suggest that RitR is an important response regulator whose primary role is to maintain iron homeostasis inS. pneumoniae. The nameritR(repressor of iron transport) for the orphan response regulator gene,rr489, is proposed.