A stm0551 knockout mutant strain constructed in the present study enabled it to produce type 1 fimbriae on the solid LB agar medium. This phenotype was correlated with the RT-PCR result that the mRNA expression of the major fimbrial subunit, fimA, was enhanced on solid-agar culture medium. These suggested that stm0551 plays a repressive role in type 1 fimbrial regulation perhaps in a similar Regorafenib manner to the role played by FimW in the fim regulatory circuit
. The expression of fimA of the transformant Δstm0551 (pSTM0551) grown on agar decreased to the same level as that of the parental LB5010 strain grown in the same conditions. However, this transformant did not exhibit visible yeast agglutination and guinea pig erythrocyte
hemagglutination when grown in static broth, nor did this strain exhibit fimA expression, which was unexpected. One https://www.selleckchem.com/products/nec-1s-7-cl-o-nec1.html of the reasons could have been the relatively high level of STM0551 production due to presence of the multiple SU5402 chemical structure copies of the pSTM0551 recombinant plasmid in these cells. An excessive STM0551 level in S. Typhimurium could presumably cause a dramatically decreased concentration of c-di-GMP locally, and subsequently interfere with fimA expression. However, the mechanism by which STM0551 interacts with fimA gene expression remains unclear. One possibility is that the stm0551 product maintained the local concentration of c-di-GMP at a level such that only a certain amount of c-di-GMP was bound by a hypothetical PilZ domain containing protein. This low concentration of c-di-GMP-bound, PilZ domain-containing protein was not able to activate fimA gene expression. Disruption of stm0551 increased the local c-di-GMP concentration and consequently Astemizole also increased the “functional” PilZ domain-containing protein to enhance fimA expression. The FimY protein of S. Typhimurium could possibly function as such a PilZ domain-containing protein since recently we found that the amino acid sequence of FimY demonstrated relatedness to those of MrkH of K. pneumoniae and YcgR of the E. coli K-12 strain (data not shown). Both MrkH and YcgR were shown to
be transcriptional activators with c-di-GMP-binding PilZ domains [28, 29]. Our hypothesis about the role FimY correlates with the finding that STM0551 did not affect fimY at the transcriptional level (Figure 5, panel C). More detailed study of FimY is necessary to define its role in a possible c-di-GMP regulatory network. Both FimY and FimZ are required to activate fimA expression in S. Typhimurium . FimZ is a DNA binding protein that binds the fimA promoter and activate its expression . Our qRT-PCR results demonstrated very similar profiles for both fimA and fimZ expression (Figure 5, panel A and B). According to the results reported by Saini et al., FimY and FimZ independently activate the fimA gene expression, in addition, FimY and FimZ also activated each other’s expression .