A previously uncharacterized gene stm0551 plays a repressive role in the regulation of type 1 fimbriae in Salmonella enterica serotype Typhimurium

Abstract

BackgroundSalmonella enterica serotype Typhimurium produces surface-associated fimbriae that facilitate adherence of the bacteria to a variety of cells and tissues. Type 1 fimbriae with binding specificity to mannose residues are the most commonly found fimbrial type. In vitro, static-broth culture favors the growth of S. Typhimurium with type 1 fimbriae, whereas non-type 1 fimbriate bacteria are obtained by culture on solid-agar media. Previous studies demonstrated that the phenotypic expression of type 1 fimbriae is the result of the interaction and cooperation of the regulatory genes fimZ, fimY, fimW, and fimU within the fim gene cluster. Genome sequencing revealed a novel gene, stm0551, located between fimY and fimW that encodes an 11.4-kDa putative phosphodiesterase specific for the bacterial second messenger cyclic-diguanylate monophosphate (c-di-GMP). The role of stm0551 in the regulation of type 1 fimbriae in S. Typhimurium remains unclear.ResultsA stm0551-deleted stain constructed by allelic exchange constitutively produced type 1 fimbriae in both static-broth and solid-agar medium conditions. Quantative RT-PCR revealed that expression of the fimbrial major subunit gene, fimA, and one of the regulatory genes, fimZ, were comparably increased in the stm0551-deleted strain compared with those of the parental strain when grown on the solid-agar medium, a condition that normally inhibits expression of type 1 fimbriae. Following transformation with a plasmid possessing the coding sequence of stm0551, expression of fimA and fimZ decreased in the stm0551 mutant strain in both culture conditions, whereas transformation with the control vector pACYC184 relieved this repression. A purified STM0551 protein exhibited a phosphodiesterase activity in vitro while a point mutation in the putative EAL domain, substituting glutamic acid (E) with alanine (A), of STM0551 or a FimY protein abolished this activity.ConclusionsThe finding that the stm0551 gene plays a negative regulatory role in the regulation of type 1 fimbriae in S. Typhimurium has not been reported previously. The possibility that degradation of c-di-GMP is a key step in the regulation of type 1 fimbriae warrants further investigation.

Dissecting the cyclic di‐guanylate monophosphate signalling network regulating motility in Salmonella enterica serovar Typhimurium

“…Typhimurium (Clegg and Hughes, 2002). Interestingly, FimZ and type 1 fimbriae expression are negatively regulated by the EAL-like protein STM0551 (Wang et al, 2012). We have previously shown that type 1 fimbriae-mediated biofilm formation temporarily precedes CsgD-mediated biofilm formation (Monteiro et al, 2012).…”

Arginine within a specific motif near the N‐terminal of FimY is critical for the maximal production of type 1 fimbriae in Salmonella enterica serovar Typhimurium

“…Within the elements of the fim gene cluster of S . Typhimurium only stm0551 had been described to involve in c‐di‐GMP metabolism, whose product is a phosphodiesterase to degrade c‐di‐GMP (Wang et al, ). No PilZ domain protein had been reported previously.…”

Salmonella Fimbriae: What is the Clue to Their Hairdo?

“…In Salmonella, curli expression was activated by AdrA, a GGDEF-domain protein that increases intracellular level of cyclic-di-GMP [61]. Fimbrial production regulated by the cyclic-di-GMP level was also observed in other species such as Klebsiella pneumonia, E. coli and P. aeruginosa [62].…”

“…The fimC and fimD genes encode respectively the chaperone and the usher [67]. Ancillary genes, fimZYW, and a rare arginine transfer RNA (tRNA) fimU and STM0551, inserted between fimY and fimW, directly regulate fim expression [62].…”

“…At the opposite, FimW repressed directly fimA expression and indirectly by lowering FimZ availability by its degradation caused by FimW binding [74]. It is also suggested that STM0551, an EAL domain protein, is a negative regulator of fim expression by lowering the c-di-GMP level [62].…”