Investigating the role of BN-domains of FlhF involved in flagellar synthesis in Campylobacter jejuni

Li, Xiaofei; Chai, Qinwen; Zheng, Lina; Huang, Pingyu; Gundogdu, Ozan; Jiao, Xinan; Tang, Yuanyue; Huang, Jinlin

doi:10.1016/j.micres.2021.126944

Cited by 4 publications

(5 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a conclusion, the GDP-bound and GTP-bound states of FlhF can be distinguished by the topology of their N domains. As this domain has been shown to affect correct flagellar assembly, an as yet unexplored signaling mechanism can be hypothesized (Li et al, 2022).…”

Section: Resultsmentioning

confidence: 99%

Structure of the GDP-bound state of the SRP GTPase FlhF

Dornes,

Mais,

Bange

2024

Acta Cryst Sect F

View full text Add to dashboard Cite

The GTPase FlhF, a signal recognition particle (SRP)-type enzyme, is pivotal for spatial–numerical control and bacterial flagella assembly across diverse species, including pathogens. This study presents the X-ray structure of FlhF in its GDP-bound state at a resolution of 2.28 Å. The structure exhibits the classical N- and G-domain fold, consistent with related SRP GTPases such as Ffh and FtsY. Comparative analysis with GTP-loaded FlhF elucidates the conformational changes associated with GTP hydrolysis. These topological reconfigurations are similarly evident in Ffh and FtsY, and play a pivotal role in regulating the functions of these hydrolases.

show abstract

Section: Resultsmentioning

confidence: 99%

Structure of the GDP-bound state of the SRP GTPase FlhF

Dornes,

Mais,

Bange

2024

Acta Cryst Sect F

View full text Add to dashboard Cite

show abstract

“…In addition, FlhF indirectly stimulates Class III transcription by enabling FlgM secretion. A negative effect of flhF deletion on late flagellar gene expression has been documented in P. aeruginosa (Murray & Kazmierczak, 2006), V. cholerae (Correa et al ., 2005) V. vulnificus (Kim et al ., 2012), Campylobacter jejuni (Ren et al ., 2018; Li et al ., 2022), Helicobacter pylori (Niehus et al ., 2004), but the mechanisms behind this phenomenon have not been explored.…”

Section: Discussionmentioning

confidence: 99%

“…Consistent with its inability to evict FlgM from the cells, the Δ flhF mutant also displays reduced expression of at least three Class III (i.e., FliA-dependent) promoters (Fig 6B) . A negative effect of flhF deletion on late flagellar gene expression has been documented in P. aeruginosa (Murray & Kazmierczak, 2006), V. cholerae (Correa et al ., 2005) V. vulnificus (Kim et al ., 2012), Campylobacter jejuni (Ren et al ., 2018; Li et al ., 2022), Helicobacter pylori (Niehus et al ., 2004), but the mechanisms behind this phenomenon have not been explored. We conclude that, in addition to its function in the determination of flagellar location, FlhF is necessary for efficient flagellar assembly beyond completion of the fT3SS.…”

Section: Discussionmentioning

confidence: 99%

Spatial, temporal and numerical regulation of polar flagella assembly inPseudomonas putida

Pulido-Sánchez,

Leal-Morales,

López-Sánchez

et al. 2023

Preprint

View full text Add to dashboard Cite

The Gram-negative bacteriumPseudomonas putidabears a tuft of flagella at a single cell pole. New flagella must be assembledde novoevery cell cycle to secure motility of both daughter cells. Here we show that the coordinated action of FimV, FlhF and FleN sets the location, timing and number of flagella assembled. The polar landmark protein FimV recruits FleN and enables stable association of FlhF to the cell poles. FlhF determines flagellar position by recruiting FliF to the cell pole and preventing its nucleation at non-polar sites. FlhF is also required for assembly of the flagellar type III secretion system, enabling Class III flagellar promoter activation and export of the external flagellar components. FleN regulates flagellar number by repressingflhFexpression and a second, posttranscriptional mechanism. Flagellar assembly starts with FlhF recruitment to the new pole shortly after cell division and finishes with the emergence of FliC filaments during the following cell division. FimV and FleN regulate the onset of flagellar assembly by preventing premature polar targeting of FlhF. Our results shed new light on the mechanisms that ensure the timely assembly of the appropriate number of flagella at the correct polar location in polarly flagellated bacteria.

show abstract

“…TEM analysis was performed as previously described with modifications [ 14 ]. One hundred µL phage stock was dropped onto a mica sheet (~3 × 3 mm size) and absorbed for 10 min.…”

Section: Methodsmentioning

confidence: 99%

“…Δ motA , Δ kpsM , Δ flaAB , Δ 06810, and Δ 06875 mutants were constructed in C. jejuni NCTC12662 by replacing target genes with aphA encoding the kanamycin-resistant cassette or cat encoding chloramphenicol resistance as previously described ( Table S1 ) [ 14 ]. In brief, the up and down flanking regions of motA , kpsM , flaAB , 06810 , 06875, and kanamycin resistant gene aph were amplified by PCR using the primers list in Table S3 .…”

Section: Methodsmentioning

confidence: 99%

Campylobacter jejuni Developed the Resistance to Bacteriophage CP39 by Phase Variable Expression of 06875 Encoding the CGPTase

Tang

YueXuan

et al. 2022

Viruses

Self Cite

View full text Add to dashboard Cite

Bacteriophage (phage) is regarded as an antimicrobial alternative for Campylobacter in food production. However, the development of phage resistance to the host is a main concern for the phage application. This study characterized the phage CP39 and investigated the phage resistance of CP39 in Campylobacter jejuni NCTC12662. We determined that phage CP39 belonged to the Myoviridae family by the WGS and phylogenetic analysis. Phage CP39 was confirmed as a capsular polysaccharide (CPS)-dependent phage by primary C. jejuni phage typing. It was further confirmed that the phage could not be adsorbed by the acapsular mutant ΔkpsM but showed the same lytic ability in both the wild-type strain NCTC 12662 and the ΔmotA mutant lacking motile flagella filaments. We further determined that the 06875 gene encoding CDP-glycerol:poly (glycerophosphate) glycerophosphotransferase (CGPTase) in the CPS loci was related to phage CP39 adsorption by SNP analysis and observed a rapid development of phage resistance in NCTC 12662 during the phage infection. Furthermore, we observed a high mutation frequency of 06875 (32%), which randomly occurred in nine different sites in the gene according to colony PCR sequencing. The mutation of the 06875 gene could cause the phase variable expression of non-functional protein and allow the bacteria against the phage infection by modifying the CPS. Our study confirmed the 06875 gene responsible for the CPS-phage adsorption for the first time and demonstrated the phase variable expression as a main mechanism for the bacteria to defend phage CP39. Our study provided knowledge for the evolutionary adaption of bacteria against the bacteriophage, which could add more information to understand the phage resistance mechanism before applying in the industry.

show abstract

Investigating the role of BN-domains of FlhF involved in flagellar synthesis in Campylobacter jejuni

Cited by 4 publications

References 32 publications

Structure of the GDP-bound state of the SRP GTPase FlhF

Structure of the GDP-bound state of the SRP GTPase FlhF

Spatial, temporal and numerical regulation of polar flagella assembly inPseudomonas putida

Campylobacter jejuni Developed the Resistance to Bacteriophage CP39 by Phase Variable Expression of 06875 Encoding the CGPTase

Contact Info

Product

Resources

About