Molecular analysis of a Leptospira borgpetersenii gene encoding an endoflagellar subunit protein

Mitchison, Marina; Rood, Julian I.; Faine, S.; Adler, Ben

doi:10.1099/00221287-137-7-1529

Cited by 29 publications

(21 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The experiments reported here exploit the use of electroporation and allelic-exchange mutagenesis as a means to better understand the function of specific PF genes in B. hyodysenteriae. Because the structure and composition of the PFs of B. hyodysenteriae (13,(25)(26)(27) are so similar to those of T. denticola (47), L. interrogans (37,52), S. aurantia (3,4,8,41), and the more intractable T. pallidum (39,40), the results obtained are likely to be relevant to these other species.…”

Section: Discussionmentioning

confidence: 86%

The Spirochete FlaA Periplasmic Flagellar Sheath Protein Impacts Flagellar Helicity

et al. 2000

View full text Add to dashboard Cite

Spirochete periplasmic flagella (PFs), including those from Brachyspira (Serpulina), Spirochaeta, Treponema, and Leptospira spp., have a unique structure. In most spirochete species, the periplasmic flagellar filaments consist of a core of at least three proteins (FlaB1, FlaB2, and FlaB3) and a sheath protein (FlaA). Each of these proteins is encoded by a separate gene. Using Brachyspira hyodysenteriae as a model system for analyzing PF function by allelic exchange mutagenesis, we analyzed purified PFs from previously constructed flaA::cat, flaA::kan, and flaB1::kan mutants and newly constructed flaB2::cat and flaB3::cat mutants. We investigated whether any of these mutants had a loss of motility and altered PF structure. As formerly found with flaA::cat, flaA::kan, and flaB1::kan mutants, flaB2::cat and flaB3::cat mutants were still motile, but all were less motile than the wild-type strain, using a swarm-plate assay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis indicated that each mutation resulted in the specific loss of the cognate gene product in the assembled purified PFs. Consistent with these results, Northern blot analysis indicated that each flagellar filament gene was monocistronic. In contrast to previous results that analyzed PFs attached to disrupted cells, purified PFs from a flaA::cat mutant were significantly thinner (19.6 nm) than those of the wild-type strain and flaB1::kan, flaB2::cat, and flaB3::cat mutants (24 to 25 nm). These results provide supportive genetic evidence that FlaA forms a sheath around the FlaB core. Using high-magnification dark-field microscopy, we also found that flaA::cat and flaA::kan mutants produced PFs with a smaller helix pitch and helix diameter compared to the wild-type strain and flaB mutants. These results indicate that the interaction of FlaA with the FlaB core impacts periplasmic flagellar helical morphology.

show abstract

Section: Discussionmentioning

confidence: 86%

The Spirochete FlaA Periplasmic Flagellar Sheath Protein Impacts Flagellar Helicity

et al. 2000

View full text Add to dashboard Cite

show abstract

“…The 29-kDa protein is of the same size as a protein that copurifies with the PFs from T. pallidum (34). FlaA and FlaB PF proteins has been found in other spirochete genera, including Spirochaeta (3,36), Treponema (2, 34), Leptospira (33,47), Serpulina, and more recently Borrelia (8,12,23,27,35). The N-terminal amino acid sequences of the various Fla proteins indicate a close phylogenetic relationship of T. denticola to T. pallidum and to T. phagedenis.…”

Section: Vol 179 1997mentioning

confidence: 97%

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

et al. 1997

View full text Add to dashboard Cite

Treponema denticola is an anaerobic, motile, oral spirochete associated with periodontal disease. We found that the periplasmic flagella (PFs), which are located between the outer membrane sheath and cell cylinder, influence its morphology in a unique manner. In addition, the protein composition of the PFs was found to be quite complex and similar to those of other spirochetes. Dark-field microscopy revealed that most wild-type cells had an irregular twisted morphology, with both planar and helical regions, and a minority of cells had a regular right-handed helical shape. High-voltage electron microscopy indicated that the PFs, especially in those regions of the cell which were planar, wrapped around the cell body axis in a right-handed sense. In those regions of the cell which were helical or irregular, the PFs tended to lie along the cell axis. The PFs caused the cell to form the irregular shape, as two nonmotile, PF-deficient mutants (JR1 and HL51) were no longer irregular but were right-handed helices. JR1 was isolated as a spontaneously occurring nonmotile mutant, and HL51 was isolated as a site-directed mutant in the flagellar hook gene flgE. Consistent with these results is the finding that wild-type cells with their outer membrane sheath removed were also right-handed helices similar in shape to JR1 and HL51. Purified PFs were analyzed by two-dimensional gel electrophoresis, and several protein species were identified. Western blot analysis using antisera to Treponema pallidum PF proteins along with N-terminal amino acid sequence analysis indicated T. denticola PFs are composed of one class A sheath protein of 38 kDa (FlaA) and three class B proteins of 35 kDa (FlaB1 and FlaB2) and one of 34 kDa (FlaB3). The N-terminal amino acid sequences of the FlaA and FlaB proteins of T. denticola were most similar to those of T. pallidum and Treponema phagedenis. Because these proteins were present in markedly reduced amounts or were absent in HL51, PF synthesis is likely to be regulated in a hierarchy similar to that found for flagellar synthesis in other bacteria.Spirochetes are recognized for their unique cell morphology and unusual means of motility (5, 48). In most spirochetes, the primary structural component of the cell body is a flexible yet semirigid helically shaped cell cylinder (5). Borrelia burgdorferi is an exception, since it has a rod-shaped cell cylinder (13, 15). Helical periplasmic flagella (PFs) are closely associated with the spirochetal protoplasmic cell cylinder (PC) and are embedded in the cytoplasmic membrane and cell wall near each end of the cell (5, 6, 17); each PF is inserted at only one end. Surrounding the entire cell is an outer membrane sheath (OS) (5). Genetic evidence indicates that the PFs are essential for the motility of several spirochete species (4,8,26,28,37,44). The PFs vary in number, length, and protein composition among the spirochete species. Depending on the species, the PFs may or may not overlap in the center of the cell (5, 8). PFs from most spirochete species are co...

show abstract

“…The promoter sequences of the Treponema fzaA genes are similar to the consensus 0 " promoter, while thefzaB genes appear to use a a28-like promoter as for other eubacterial motility genes (Table 2). Recently, the flaB gene of Leptospira borgpetersenii and the fzaB2 gene of Serptllina (Treponema) h_yo&senteriae were sequenced and no promoter sequences were found (Mitchison et al, 1991;Koopman et al, 1993). No transcriptional start point has been determined for these genes, but when we examined the flaB 5'-noncoding region of L. borgpetersenii and S. byo&enteriae, we discovered a putative promoter sequence, 82 and 65 bp, respectively, upstream of the ATG start codon.…”

Section: Ctcaa Gtgcacccggcgtatt Ccgatac Ctcaa G T C C T T T T C C G Tmentioning

confidence: 99%

Expression of the flagellin gene in Borrelia is controlled by an alternative factor

Noppa

Burman²,

Sadziene³

et al. 1995

Microbiology

View full text Add to dashboard Cite

show abstract

Molecular analysis of a Leptospira borgpetersenii gene encoding an endoflagellar subunit protein

Cited by 29 publications

References 39 publications

The Spirochete FlaA Periplasmic Flagellar Sheath Protein Impacts Flagellar Helicity

The Spirochete FlaA Periplasmic Flagellar Sheath Protein Impacts Flagellar Helicity

Relationship of Treponema denticola periplasmic flagella to irregular cell morphology

Expression of the flagellin gene in Borrelia is controlled by an alternative factor

Contact Info

Product

Resources

About