AUTHOR=Hirakawa Hidetada , Shimokawa Mizuki , Noguchi Koshi , Tago Minori , Matsuda Hiroshi , Takita Ayako , Suzue Kazutomo , Tajima Hirotaka , Kawagishi Ikuro , Tomita Haruyoshi 

TITLE=The PapB/FocB family protein TosR acts as a positive regulator of flagellar expression and is required for optimal virulence of uropathogenic Escherichia coli

JOURNAL=Frontiers in Microbiology

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2023.1185804

DOI=10.3389/fmicb.2023.1185804

ISSN=1664-302X

ABSTRACT=<p>Uropathogenic <italic>Escherichia coli</italic> (UPEC) is a major causative agent of urinary tract infections. The bacteria internalize into the uroepithelial cells, where aggregate and form microcolonies. UPEC fimbriae and flagella are important for the formation of microcolonies in uroepithelial cells. PapB/FocB family proteins are small DNA-binding transcriptional regulators consisting of approximately 100 amino acids that have been reported to regulate the expression of various fimbriae, including P, F1C, and type 1 fimbriae, and adhesins. In this study, we show that TosR, a member of the PapB/FocB family is the activator of flagellar expression. The <italic>tosR</italic> mutant had similar expression levels of type 1, P and F1C fimbriae as the parent strain, but flagellar production was markedly lower than in the parent strain. Flagellin is a major component of flagella. The gene encoding flagellin, <italic>fliC</italic>, is transcriptionally activated by the sigma factor FliA. The <italic>fliA</italic> expression is induced by the flagellar master regulator FlhDC. The <italic>flhD</italic> and <italic>flhC</italic> genes form an operon. The promoter activity of <italic>fliC, fliA</italic> and <italic>flhD</italic> in the <italic>tosR</italic> mutant was significantly lower than in the parent strain. The purified recombinant TosR does not bind to <italic>fliC</italic> and <italic>fliA</italic> but to the upstream region of the <italic>flhD</italic> gene. TosR is known to bind to an AT-rich DNA sequence consisting of 29 nucleotides. The characteristic AT-rich sequence exists 550–578 bases upstream of the <italic>flhD</italic> gene. The DNA fragment lacking this sequence did not bind TosR. Furthermore, loss of the <italic>tosR</italic> gene reduced motility and the aggregation ability of UPEC in urothelial cells. These results indicate that TosR is a transcriptional activator that increases expression of the <italic>flhDC</italic> operon genes, contributing to flagellar expression and optimal virulence.</p>