The UzcRS two-component system integrates regulatory input from a diverse set of auxiliary regulators
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ABSTRACT:
PROVIDER: PRJNA422404 | ENA |
REPOSITORIES: ENA
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Project description:The UzcRS two-component system in Caulobacter crescentus functions as a global activator of an extracytoplasmic stress response pathway and is induced by the metals U, Zn and Cu. Unexpectedly, a screen for mutations that affected the activity of the UzcR-regulated urcA promoter (PurcA) revealed four previously uncharacterized proteins whose inactivation led to metal-independent induction of PurcA. Using molecular genetics and functional genomics, we found that these auxiliary regulators regulate PurcA expression by modulating the activity of UzcRS through distinct mechanisms. The membrane protein UzcX, whose expression is positively regulated by UzcR, forms a regulatory complex with UzcS, providing a mechanism of feedback inhibition within the UzcRS circuit. UrtAP, an ABC transporter with a periplasmic metallo-aminopeptidase domain, negatively regulates UzcRS activity and antagonizes metal dependent stimulation by virtue of its enzymatic activity. Lastly, two MarR family transcription factors, MarR1 and MarR2, synergistically regulate UzcRS activity by repressing the expression of the membrane proteins UzcY and UzcZ, respectively, which function as UzcRS activators. In addition to modulating basal UzcRS activity, UzcY and UzcZ enhance the U/Zn/Cu sensitivity of UzcRS, shifting its responsiveness to more environmentally-relevant metal concentrations. Collectively, these data suggest that UzcRS functions as the core-signaling unit within a multicomponent signal transduction pathway that includes an unusually diverse set of auxiliary regulators.
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Project description:In response to osmolarity, Salmonella enterica serotype Typhi (S. Typhi) regulates genes required for Vi capsular antigen expression oppositely to those required for motility and invasion. Previous studies suggest that osmoregulation of motility, invasion and capsule expression is mediated through the RcsC/RcsD/RcsB phosphorelay system. Here we performed gene expression profiling and functional studies to determine the role of TviA, an auxiliary protein of the RcsB response regulator, in controlling virulence gene expression in S. Typhi. TviA repressed expression of genes encoding flagella and the invasion associated type III secretion system (T3SS-1) through repression of the flagellar regulators flhDC and fliZ, resulting in reduced invasion and reduced expression of FliC. Both RcsB and TviA reduced expression of flhDC, but only TviA altered flhDC expression in response to osmolarity. These data suggest that the auxiliary TviA protein integrates a new regulatory input into the RcsB regulon of S. Typhi, thereby altering expression of genes encoding flagella, the Vi antigen and T3SS-1 in response to osmolarity. Gene expression profiles of the S. Typhi wild-type strain, a rcsB mutant, a (delta)tviB-vexE (tviBCDEvexABCDE) mutant, and a (delta)viaB (tviABCDEvexABCDE) mutant (lacking tviA expression) was compared. Strains were cultured in SOB broth (low salt), a condition known to induce expression of TviA and the RcsCB system. Total RNA was extracted from two independent cultures (biological replicates) of each strain. cDNA was differentially labeled with Alexa Fluor 555 and 648, respectively and competitively hybridized on S.Typhimurium arrays (PFGRC, version 4). The submitted samples are biological replicates of a competitive hybridization of cDNA from the wild-type strain (Ty2) and the rcsB mutant (SW237) (GSM395580, GSM395581) or the viaB mutant (STY2) and the tviB-vexE mutant (SW74) (GSM395582, GSM395583), respectively.
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