the response regulator CsgD in turn regulates the transcription of adrA
In order to determine which CsgD-regulated factor , either curli fimbriae or cellulose , was responsible for restoring the biofilm phenotype in 3934 ∆ stm2689 [ pBR328 : : csgD ] , 3934 ∆ stm2689 strain was complemented with the adrA gene under the control of its own promoter , which exclusively activates cellulose A C Arabinose 0.1 % Glucose 0.1 % 3934 3934BADstm2689 B 3934BADstm2689 3934 3934 ara gluc ∆ stm2689 205 kD Fig. 3 .
In order to determine which CsgD-regulated factor , either curli fimbriae or cellulose , was responsible for restoring : csgD , 3934 ∆ stm2689 strain was complemented with the adrA gene under the control of of its own prom .
In order to determine which CsgD-regulated factor , either curli fimbriae or cellulose , was responsible for restoring the biofilm phenotype in 3934 ∆ stm2689 [ pBR328 , 3934 ∆ stm2689 strain was complemented with the adrA gene under the control of of its own prom .
Altogether , these results show that bapA , like adrA , is regulated by CsgD , which plays a central role in biofilm formation in Salmonella strains .
Positive regulation by CsgD is indicated by solid lines because putative CsgD binding sites have been found upstream from the adrA promoters .
CsgD controls the expression of adrA .
Indeed , it has been suggested , although it has never been shown , that CsgD binds to conserved 11-bp sequences upstream of the adrA promoters , activating transcription of these genes .
Results CsgD binds directly to adrA promoter regions In S. Typhimurium the expression of adrA is positively regulated by the transcriptional regulator CsgD .
Results CsgD binds directly to adrA promoter regions In S. Typhimurium the expression of adrA is positively regulated by the transcriptional regulator CsgD .
Results CsgD binds directly to adrA promoter regions In S. Typhimurium the expression of csgBA is positively regulated by the transcriptional regulator CsgD .
Results CsgD binds directly to the csgBA In S. Typhimurium the expression of adrA is positively regulated by the transcriptional regulator CsgD .
In order to demonstrate the direct binding of CsgD to the adrA promoter regions we performed EMSA with purified CsgD-His6 tagged protein ( Fig .
In order to demonstrate the direct binding of CsgD to the adrA promoter regions we performed electrophoretic-mobility-shift assays with purified CsgD-His6 tagged protein ( Fig .
Since CsgD is highly integrated into the c-di-GMP signalling network , we investigated whether c-di-GMP affects the binding of CsgD-His6 to adrA promoter region .
To investigate the binding of CsgD to the adrA promoter region , a 257 bp 32P-labelled fragment from bp -222 to +35 was subjected to Fig. 2E .
To investigate the binding of CsgD to the adrA promoter region , a 257 bp 32P-labelled fragment from bp -222 to +35 was subjected to DNase I footprint analysis .
Pre-incubation of CsgD-His6 with 10 mM AcP led to a decreased binding of CsgD to the adrA , compared with Fig. 5A .
Pre-incubation of CsgD-His6 with 10 mM AcP led to a decreased binding of CsgD to the adrA , compared with CsgD alone .
Since the binding of CsgD to the adrA promoter regions is altered in the presence of the phosphodonor AcP , we investigated whether CsgD can be phosphorylated in-vitro .
However , we discovered that c-di-GMP , is not required for binding of CsgD to the adrA promoters .
However , we discovered that an integral part of the regulatory network , is not required for binding of CsgD to the adrA promoters .
Typhimurium cells _ treated with polyphenol , suggesting that the CsgD can regulate the transcription of adrA gene involved in bio-film formation , EPS production
Typhimurium cells _ treated with polyphenol , suggesting that the CsgD can regulate the transcription of adrA gene involved in bio-film formation , transport production
Typhimurium cells _ treated with polyphenol , suggesting that the CsgD can regulate the transcription of adrA gene involved in bio-film formation , virulence production
Typhimurium cells _ treated with polyphenol , suggesting that the CsgD can regulate the transcription of adrA gene the upregulation of expression of lpfE genes
Typhimurium cells _ treated with polyphenol , suggesting that the CsgD can regulate the transcription of adrA gene the upregulation of expression of fimA genes
In contrast to the CsgD homolog in Vibrio cholerae , CsgD does not bind c-di-GMP as CsgD-mediated transcriptional activation of adrA is unaffected by c-di-GMP in-vitro .
In contrast to VpsT , CsgD does not bind c-di-GMP as CsgD-mediated transcriptional activation of adrA is unaffected by c-di-GMP in-vitro .
To quantitate the degree of CsgD complementation in each strain , we measured the expression of csgB , coding for adrA , coding for a regulator of cellulose production , using promoter-lux operon fusion plasmids .
To quantitate the degree of CsgD complementation in each strain , we measured the expression of csgD , coding for adrA , coding for a regulator of cellulose production , using promoter-lux operon fusion plasmids .