Twin RNA Polymerase–Associated Proteins Control Virulence Gene Expression in Francisella tularensis

PLoS Pathogens

Eric J Rubin¹  Emmy L Balon¹  Dana H Boyd²  James C Charity³  Simon L Dove³  Michelle M Costante-Hamm³ 

[1] Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America;Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts, United States of America;Division of Infectious Diseases, Children's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America

关键词: Gene expression;    Francisella tularensis;    Gene regulation;    Plasmid construction;    DNA-binding proteins;    Polymerase chain reaction;    RNA synthesis;    DNA transcription;   

DOI  :  10.1371/journal.ppat.0030084

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

The MglA 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 MglA-like protein called SspA. Here, we show that MglA and SspA cooperate with one another to control virulence gene expression in F. tularensis. Using a directed proteomic approach, we show that both MglA and SspA associate with RNA polymerase (RNAP) in F. tularensis, and that SspA is required for MglA to associate with RNAP. Furthermore, bacterial two-hybrid and biochemical assays indicate that MglA and SspA interact with one another directly. Finally, through genome-wide expression analyses, we demonstrate that MglA and SspA regulate the same set of genes. Our results suggest that a complex involving both MglA 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 α subunits of RNAP, and we show here that these two α 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.

【 授权许可】

CC BY