Flagellar Synthesis in Salmonella typhimurium: Factors Affecting the Formation of the Flagellar  -N-Methyllysine

Kerridge, D.

doi:10.1099/00221287-42-1-71

Cited by 15 publications

(9 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Alternatively, it could be that FlaC and FlaD originated from the same gene but FlaD is a posttranslationally modified FlaC. Posttranslational modification has been reported to occur in several other bacterial species (18,21,22,26,46). However, support for the likelihood of four flagellin genes may come from a V. parahaemolyticus study (29).…”

Section: Discussionmentioning

confidence: 99%

Flagellin A is essential for the virulence of Vibrio anguillarum

et al. 1996

View full text Add to dashboard Cite

A flagellin gene from the fish pathogen Vibrio anguillarum was cloned, sequenced, and mutagenized. The DNA sequence suggests that the flaA gene encodes a 40.1-kDa protein and is a single transcriptional unit. A polar mutation and four in-frame deletion mutations (180 bp deleted from the 5 end of the gene, 153 bp deleted from the 3 end of the gene, a double deletion of both the 180-and 153-bp deletions, and 942 bp deleted from the entire gene) were made. Compared with the wild type, all mutants were partially motile, and a shortening of the flagellum was seen by electron microscopy. Wild-type phenotypes were regained when the mutations were transcomplemented with the flaA gene. Protein analysis indicated that the flaA gene corresponds to a 40-kDa protein and that the flagellum consists of three additional flagellin proteins with molecular masses of 41, 42, and 45 kDa. N-terminal sequence analysis confirmed that the additional proteins were flagellins with N termini that are 82 to 88% identical to the N terminus of FlaA. Virulence studies showed that the N terminal deletion, the double deletion, and the 942-bp deletion increased the 50% lethal dose between 70-and 700-fold via immersion infection, whereas infection via intraperitoneal injection showed no loss in virulence. In contrast, the polar mutant and the carboxy-terminal deletion mutant showed approximately a 10 4 -fold increase in the 50% lethal dose by both immersion and intraperitoneal infection. In summary, FlaA is needed for crossing the fish integument and may play a role in virulence after invasion of the host.Vibrio anguillarum, which causes vibriosis in marine fish, is a highly pathogenic bacterium and has become a severe problem for the fish farming industry. An earlier study (3) has correlated the virulence of V. anguillarum with the possession of more than one flagellum. The importance of the flagellum as a potential virulence factor has been demonstrated for other bacteria. In Pseudomonas aeruginosa (5), three wild-type strains and their respective isogenic motility mutants were tested in the burned mouse model. The nonmotile mutants proliferated in the wound but did not cause the characteristic systemic infection, indicating that motility contributes to the invasive capabilities of this organism. For Campylobacter jejuni, the flagellum is the best-characterized virulence factor. In vitro studies using epithelial cell lines have shown that the flagellum aids the bacterium either in adherence to (30) or in internalization within (11) the cultured cells. A more recent study (47) shows that motility and not flagellin A is required for the invasion of intestinal cells by C. jejuni. However, flagellin A can serve as a secondary adhesin for the adherence to intestinal cells, whereas other adhesins are present to aid the motilitydependent internalization in an intestinal cell line. Similarly, for Vibrio cholerae, studies using spontaneous mutants or chemically derived motility mutants indicate that either motility (12) or an adhesin, proposed to be asso...

show abstract

Section: Discussionmentioning

confidence: 99%

Flagellin A is essential for the virulence of Vibrio anguillarum

et al. 1996

View full text Add to dashboard Cite

show abstract

“…Such posttranslational modification of bacterial flagellins has been documented in several bacterial species: specific lysines are modified to N-methyllysines in certain serotypes of S. typhimurium (19,21), while Campylobacter coli flagellin contains phosphorylated serines (30), and H. halobium produces flagellins that are sulfated glycoproteins (52). Determination of whether the P. aeruginosa flagellin can undergo modification of the amino acid side chains will require isolation of products from both bacteria as well as physical and chemical analysis.…”

Section: Gcgctccgagwgaccgct Tgaacggcgaag'jcgaagc''aactgc'gaaaga Ggmentioning

confidence: 99%

Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK

1990

View full text Add to dashboard Cite

Flagella in procaryotes are complex structures requiring the coordinate expression of over 50 genes, including flagellin, the major repeating structural protein. We have previously shown that a functional RpoN gene product is required for expression of flagellin in Pseudomonas aeruginosa PAK (P. A. Totten and S. Lory, J. Bacteriol. 172:389-396, 1990) and have now cloned, sequenced, and determined the transcriptional start site of the structural gene for this flagellin. The clones containing this gene produced a protein that reacted on Western immunoblots with polyclonal and four different monoclonal antibodies to purified flagella. However, this flagellin protein in Escherichia coli was slightly smaller (41 kDa) than flagellin protein produced in P. aeruginosa PAK (45 kDa), indicating degradation in E. coli or modification in P. aeruginosa. Comparison of the deduced amino acid sequence of this gene with the amino acid sequences of other flagellins revealed a conservation in the N-and C-terminal domains, suggesting conservation of secretion or assembly signals between these organisms. The sequence 5' of the structural gene contained potential RpoN-specific promoters as well as a promoter sequence recognized by RpoF (if28), the alternative sigma factor required for expression of flagellin genes in E. coli (and Bacillus subtilis). Deletion analysis of the promoter region as well as transcriptional start site mapping implicated the RpoF, and not the RpoN, consensus sequence as the functional promoter for the flagellin gene. Models for the involvement of both RpoN and RpoF in the expression of flagellin in P. aeruginosa are presented.Pseudomonas aeruginosa is one of the more important causative agents of opportunistic infections in humans. To cause disease, this organism needs to express an array of virulence factors, including pili, several excreted proteins and toxins, and functional polar flagella. Nonmotile variants of this organism, lacking flagella, are severely attenuated in their ability to cause disease in animal models (6). Moreover, flagella are a major antigenic determinant for the immune response to P. aeruginosa infection, and antibodies to flagella are protective (6).Flagella are found in many different genera of bacteria, and they are similar in basic structure (32). In general, flagella are anchored to the cell envelope via a basal body, which is connected to a hook structure and then extends to a long helical filament. Whereas a large number of different polypeptides make up the basal body and the hook, the flagellar filament is usually composed of one or two repeating subunits of identical polypeptide monomers, designated flagellins. The biogenesis of flagella in Escherichia coli and other bacteria involves a coordinately controlled interaction of the structural components of the basal body, hook, and filament (15,23,25 We have previously shown that in P. aeruginosa, the rpoN gene product is required for expression not only of pilin but also of flagellin (16,51). RpoN is an alternative sigma factor...

show abstract

“…Methylation of some lysine residues in the polypeptide has also been suggested by Allfrey, Faulkner & Mirsky (1964). Support for this methylation scheme has come from genetic studies (Stocker et al 1961), from an absence of an effect on methylation with addition of t-RNA, amino acid or energy source (Comb et al 1966) and from inhibition of methylation by methionine analogues although lysine incorporation was unaffected (Kerridge, 1966).…”

Section: Amino Acids Occasionally Encountered In Proteinsmentioning

confidence: 99%

Amino Acid Selection in Protein Biosynthesis

Peterson¹

1967

Biological Reviews

View full text Add to dashboard Cite

Flagellar Synthesis in Salmonella typhimurium: Factors Affecting the Formation of the Flagellar -N-Methyllysine

Cited by 15 publications

References 10 publications

Flagellin A is essential for the virulence of Vibrio anguillarum

Flagellin A is essential for the virulence of Vibrio anguillarum

Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK

Amino Acid Selection in Protein Biosynthesis

Contact Info

Product

Resources

About