Twin RNA polymerase -: Associated proteins control virulence gene expression in Francisella tularensis

The MgIA protein is the only known regulator of virulence gene expression in Francisella tularensis, yet it is unclear how it functions. F. tularensis also contains an MgIA-like protein called SspA. Here, we show that MgIA and SspA cooperate with one another to control virulence gene expression in F. tularensis. Using a directed proteomic approach, we show that both MgIA and SspA associate with RNA polymerase (RNAP) in F. tularensis, and that SspA is required for MgIA to associate with RNAP. Furthermore, bacterial two-hybrid and biochemical assays indicate that MgIA and SspA interact with one another directly. Finally, through genome-wide expression analyses, we demonstrate that MgIA and SspA regulate the same set of genes. Our results suggest that a complex involving both MgIA and SspA associates with RNAP to positively control virulence gene expression in F. tularensis. The F. tularensis genome is unusual in that it contains two genes encoding different a subunits of RNAP, and we show here that these two a subunits are incorporated into RNAP. Thus, as well as identifying SspA as a second critical regulator of virulence gene expression in F. tularensis, our findings provide a framework for understanding the mechanistic basis for virulence gene control in a bacterium whose transcription apparatus is unique.