Search Results (132)

In this study, we found that the deletion of the gene evfG in the type VI secretion system (T6SS) gene cluster significantly affected the motility of porcine extraintestinal pathogenic Escherichia coli (ExPEC) strain PCN033. Furthermore, the bacterial motility assay showed that ΔevfG mutants exhibited reduced motility compared to the parental strain. Transmission electron microscopy (TEM) showed a significant reduction in the number of flagella in the mutant ΔevfG when compared with PCN033. To further explore the reasons why the deletion of evfG affects the motility of PCN033, transcriptomic and metabolomic analyses were conducted. The omics analyses showed that 134 differentially accumulated metabolites and 2236 differentially expressed genes were identified between the mutant ΔevfG and the parental strain PCN033. The metabolome profile and functional annotation analyses indicated that the impaired motility of ΔevfG was connected to the downregulation of the expression levels of genes associated with the energy metabolism pathway and flagellar assembly. Our study provides a new insight into the diminished PCN033 motility induced by evfG deletion. Moreover, the candidate genes and metabolites regulated by the gene evfG in the T6SS, which was involved in the motility of PCN033, were reported in this study. Full article

It is well established that flagella play a critical role in bacterial motility and virulence, and the genes associated with flagellar synthesis and regulation have been extensively characterized. In this study, we identified the trmE gene as a novel modulator of flagellar synthesis in Salmonella Enteritidis. A transposon (Tn5) mutant library of Salmonella Enteritidis (SE) was constructed through bacterial conjugation, followed by screening for motility-deficient mutants. Among 1321 mutants screened, C50041trmE::Tn5 exhibited reduced motility. To validate this phenotype, we constructed C50041ΔtrmE mutants and complementary strains C50041ΔtrmE::trmE. Compared to parental strain SE(C50041), C50041ΔtrmE displayed significantly lower mRNA levels of flagellar synthesis-related genes as determined via quantitative real-time PCR and the few visible flagella observed via transmission electron microscopy (TEM). Function studies assessing virulence also showed results that matched this phenotype; specifically, C50041ΔtrmE demonstrated decreased adhesion and invasion capabilities towards macrophages. Furthermore, C50041ΔtrmE induced impaired apoptosis and pyroptosis in macrophages, while exhibiting reduced mortality in BALB/c mice along with diminished mRNA levels of pro-inflammatory cellular factors within murine spleen. This study provides compelling evidence that the trmE gene in Salmonella Enteritidis is involved in flagellar synthesis. Full article

In the study, a comprehensive investigation on potential bacterial pathogens affecting largemouth bass (Micropterus salmoides) was performed. Monthly surveys were conducted from April to October 2024. Diseased largemouth bass exhibited diverse clinical symptoms, such as rot of gill and fin, ulcers on body surface, and petechial hemorrhages in liver. Following isolation and identification, a total of 21 potential bacterial pathogens (numbered strain 1 to 21, respectively) were identified. The genus Aeromonas had the highest proportion (67.14%), among which the frequency of Aeromonas veronii was 24.60%. TEM analysis revealed that the bacterial strains exhibited three predominant shapes (rod-shaped, spherical, and curved) with length ranging from 0.5 to 3 μm. Flagellar structures were observed in strains 1–4, 6–8, 11–17, and 19–21, with variations in number and growth sites. Three isolates (strains 9, 10, 18) demonstrated Gram-positive characteristic, and strains 5, 11, and 18 have capsule structures. Strains 5, 9, 10, and 18 were non-motile, and strains 1–4, 6, 7, 9–11, 16–18, and 21 exhibited β-hemolysis. Physiological and biochemical characteristics of the 21 bacterial isolates were comprehensively analyzed. Antibiotic sensitivity testing revealed that florfenicol and enrofloxacin exhibited excellent antibacterial effects. These data will enrich the potential bacterial diseases information and promote the healthy development of the largemouth bass industry. Full article

Aeromonas hydrophila ST251 is a crucial pathogen responsible for the outbreaks of Motile Aeromonas Septicemia (MAS) in global aquaculture. To elucidate the genetic basis underlying its hypervirulence, we investigated strain D4, an ST251 isolate recovered from diseased blunt snout bream. Phenotypic assays revealed that, compared to the environmental strain ATCC 7966^T, D4 exhibited enhanced motility, hemolytic activity, and protease production. Average nucleotide identity (ANI) analysis demonstrated that D4 clustered within a distinct ST251 clade, with ANI values ≥ 99.74%. Comparative genomic analysis of D4, nine additional ST251 strains, and ATCC 7966^T identified multiple unique genomic islands in ST251 strains, including pathways for myo-inositol and L-fucose utilization and a pseudaminic acid biosynthesis gene cluster. These genetic elements are associated with nutrient acquisition and flagellar assembly, potentially enhancing colonization and environmental adaptability. In addition, distinct plasmids and prophages in ST251 strains may contribute to host adaptation and virulence by regulating stress responses and virulence-associated genes. These findings offer new insights into the molecular mechanisms driving the pathogenicity and adaptability of hypervirulent A. hydrophila ST251 strains. Full article

Extreme acidophiles from the Acidithiobacillia class thrive in highly acidic environments where they rely on diverse regulatory mechanisms for adaptation. These mechanisms include sigma factors, transcription factors (TFs), and transcription factor binding sites (TFBS), which control essential pathways. Comparative genomics and bioinformatics analyses identified sigma factors and TFs in Acidithiobacillia, showing similarities but key differences from reference neutrophiles. This study highlights sigma54-dependent one- and two-component systems that are crucial for survival in energy acquisition from sulfur compounds and hydrogen as well as nutrient assimilation. Furthermore, the data suggested evolutionary divergence in regulatory elements distinguishes S-oxidizing from Fe-S-oxidizing members of Acidithiobacillia. Conservation of gene clusters, synteny, and phylogenetic analyses supported the expected phenotypes in each species. Notable examples include HupR’s role in hydrogenase-2 oxidation in Fe-S-oxidizers, TspR/TspS regulation of the sulfur oxidation complex, and FleR/FleS control of flagellar motility in S-oxidizers. These regulatory mechanisms act as master controllers of bacterial activity, reflecting adaptation to distinct metabolic needs within Acidithiobacillia. Full article

Listeria monocytogenes is of a significant concern for the food industry, largely due to its ability to form biofilms. Flagellar motility and environmental factors are crucial for biofilm formation. Cysteine is an important compound affecting the behavior of this bacterium; therefore, we investigated its role in growth, biofilm formation and motility of L. monocytogenes 10403S through a mutant in cysteine uptake (ΔctaP). Basal defined media (DM) and L-cysteine-supplemented DM were used. Biofilm formation was promoted by L-cysteine supplementation in both wild type (WT) and ΔctaP. Lower biofilm formation of ΔctaP compared to WT indicates the significance of the cysteine transporter and cysteine uptake. A negative correlation was found between growth and biofilm formation, especially in the presence of high L-cysteine concentrations. Motility experiments showed that as the L-cysteine concentration increased, the swarming motility of WT decreased. Furthermore, swimming motility of WT was enhanced with L-cysteine supplementation, while the swimming motility of ΔctaP remained unaffected. To evaluate the role of cysteine and CtaP in biofilm formation and motility, transcriptome analysis, comparing WT and ΔctaP in basal and L-cysteine-supplemented (1.57 and 3.67 mM) DM, was conducted at 37 °C. The investigation of biofilm-related genes explained the role of ctaP and revealed induced expression of flagella and chemotaxis genes by L-cysteine. Full article

The fish pathogen Yersinia ruckeri forms biofilms on abiotic surfaces, contributing to recurrent infections in aquaculture. Increasing evidence suggests that the RNA chaperone Hfq and small non-coding RNAs (sRNAs) are key regulators of bacterial biofilm formation. However, the regulatory mechanisms mediated by these factors remain largely unexplored in Y. ruckeri. In this study, we investigated the roles of Hfq and the Hfq-dependent sRNAs RprA, ArcZ, and RybB in the biofilm formation of Y. ruckeri. We first characterized the sRNAome of biofilm-forming cells, identifying the conserved RprA, ArcZ, and RybB, among the upregulated sRNAs. We then evaluated motility, biofilm formation, and architecture in strains lacking either hfq (Δhfq) or these sRNAs (ΔsRNA). Our results reveal that both Δhfq and ΔsRNA strains exhibit significant alterations in biofilm and motility phenotypes, including changes in bacterial morphology and extracellular matrix. Furthermore, expression analyses indicate that these sRNAs modulate the transcription of key regulatory factors, flagellar and phosphodiesterase genes, ultimately influencing intracellular cyclic di-GMP levels, a key second messenger in biofilm formation. Together, our findings demonstrate that Hfq and its associated sRNAs play critical regulatory roles in Y. ruckeri biofilm formation by controlling the expression of genes involved in motility, bacterial envelope proteins, and c-di-GMP metabolism. Full article

Motile cilia play essential roles in various physiological processes including fluid flow generation and sperm motility. In this study, we identified 1,3-diphenyl-6-(4-phenylpiperazin-1-yl)benzo[e][1,2,4]triazin-7(1H)-one as a potent and reversible modulator of ciliary function using the Xenopus laevis model. This benzotriazinone derivative inhibits ciliary-driven fluid flow by inducing cilia detachment without causing toxicity in developing embryos. Unlike traditional deciliation agents that rely on calcium signaling, this compound induces cilia loss through a shear stress-driven mechanism at the transition zone, without disrupting tissue morphology or the apical actin network. Importantly, it also induces flagellar loss and impairs sperm motility at picomolar concentrations. Our findings highlight the potential of this 6-(4-phenylpiperazin-1-yl)-substituted benzotriazinone as a non-hormonal male contraceptive and underscore a novel mechanism of cilia modulation that may have broader implications for the treatment of cilia-related disorders. Full article

Bacterial pathogens employ multiple strategies to invade and damage host tissues while evading immune defenses. Recent studies highlight flagella as crucial contributors to bacterial virulence, not only by facilitating motility, but also by regulating the secretion of virulence factors. However, the role of the flagella-dependent secretome remains largely unexplored. We have recently shown that FliK, a key regulator that defines substrate specificity in the flagellar export apparatus, is essential for the resistance of Bacillus thuringiensis (B. thuringiensis) against antimicrobial peptides (AMPs) and its virulence in a Drosophila infection model. In this study, we used liquid chromatography–tandem mass spectrometry to conduct a large-scale comparative analysis of the proteins secreted in culture supernatant or associated with the cell wall of the ΔfliK mutant and its reference strain. Our results reveal significant differences in the secretome and surfaceome of the ΔfliK mutant compared to the reference strain. These findings emphasize the role of FliK in regulating the production and secretion of several proteins, underscoring the importance of flagella in controlling various biological processes. This work provides valuable insights into the functional characterization of potential candidate proteins involved in B. thuringiensis virulence and AMP resistance mechanisms. Overall, these results open new perspectives for understanding the molecular processes that govern bacterial resistance to AMPs. Full article

Vibrio parahaemolyticus, a halophilic Gram-negative bacterium commonly found in aquatic products, can colonize the human small intestine, causing gastroenteritis and potentially leukemia. As a major intestinal pathogen, it poses a significant threat to public health. This study aims to investigate the phenotypic heterogeneity of V. parahaemolyticus in the low-salinity and bile salt environments of the human intestinal tract and to elucidate its mechanisms of tolerance and pathogenicity using proteomics. The experimental results indicated that under the low salinity and bile salts conditions of the human intestinal environment, the growth, motility, and biofilm formation of the strains were significantly inhibited. Proteomics analysis revealed that, under these conditions, the energy metabolism, chemotaxis system, flagellar motor, and ribosome-related proteins of V. parahaemolyticus were significantly affected, thereby influencing its growth, motility, and biofilm formation. Furthermore, the activation of the secretion system, particularly the T2SS, enhanced the virulence of secreted factors on host cells. Additionally, the activation of the β-lactam resistance pathway increased resistance to the intestinal environment, thereby enhancing the pathogenicity of V. parahaemolyticus. Full article

In cryopreservation technology, the choice of cryoprotectant plays a crucial role in cell survival and function. Different types of cryoprotectants, each with unique protective mechanisms, mitigate cellular damage from ice crystal formation during freezing. This study investigated the effects of different types and concentrations of cryoprotectants on the cryopreservation efficacy of noble scallop Mimachlamys nobilis sperm. Six cryoprotectants were tested, including four permeable cryoprotectants (dimethyl sulfoxide (DMSO), ethylene glycerol (EG), propylene glycerol (PG), methanol (MET)) and two non-permeable cryoprotectants (trehalose (TRE), fetal bovine serum (FBS)). The results showed that permeable cryoprotectants, which penetrate the cell membrane, regulate the osmotic pressure inside and outside cells to reduce dehydration damage. Among them, 10% DMSO provided the best protection, significantly preserving sperm motility, velocity, and morphology. Non-permeable cryoprotectants, although unable to penetrate cells, stabilized the extracellular environment at higher concentrations (such as FBS). Additionally, MET and FBS exhibited enhanced protective effects with increasing concentration, indicating their potential in reducing sperm structural damage at higher concentrations. Morphological observations indicated that freezing caused varying degrees of structural damage to sperm, with flagellar integrity being crucial for motility. Overall, selecting an appropriate cryoprotectant and concentration is essential for the efficient cryopreservation of M. nobilis sperm, providing a valuable reference for conserving germplasm resources of marine species. Full article

The bacterial flagellum, a complex nanomachine composed of numerous proteins, is utilized by bacteria for swimming in various environments and plays a crucial role in their survival and infection. The flagellar motor is composed of a rotor and stator complexes, with each stator unit functioning as an ion channel that converts flow from outside of cell membrane into rotational motion. Paenibacillus sp. TCA20 was discovered in a hot spring, and a structural analysis was conducted on the stator complex using cryo-electron microscopy to elucidate its function. Two of the three structures (Classes 1 and 3) were found to have structural properties typical for other stator complexes. In contrast, in Class 2 structures, the pentamer ring of the A subunits forms a C-shape, with lauryl maltose neopentyl glycol (LMNG) bound to the periplasmic side of the interface between the A and B subunits. This interface is conserved in all stator complexes, suggesting that hydrophobic ligands and lipids can bind to this interface, a feature that could potentially be utilized in the development of novel antibiotics aimed at regulating cell motility and infection. Full article

The motility and chemotaxis of Vibrio cholerae, the bacterial pathogen responsible for cholera, play crucial roles in both environmental survival and infection. Understanding their molecular mechanisms is therefore essential not only for fundamental biology but also for infection control and therapeutic development. The bacterium’s sheathed, polar flagellum—its motility organelle—is powered by a sodium-driven motor. This motor’s rotation is regulated by the chemotaxis (Che) signaling system, with a histidine kinase, CheA, and a response regulator, CheY, serving as the central processing unit. However, V. cholerae possesses two additional, parallel Che signaling systems whose physiological functions remain unclear. Furthermore, the bacterium harbors over 40 receptors/transducers that interact with CheA homologs, forming a complex regulatory network likely adapted to diverse environmental cues. Despite significant progress, many aspects of these systems remain to be elucidated. Here, we summarize the current understanding to facilitate future research. Full article

The bacterial flagellar motor is one of the few known rotary motors, powering motility and chemotaxis. The mechanisms underlying its rotation and the switching of its rotational direction are fundamental problems in biology that are of significant interest. Recent high-resolution studies of the flagellar motor have transformed our understanding of the motor, revealing a novel gear mechanism where a membranous pentamer of MotA proteins rotates around a cell wall-anchored dimer of MotB proteins to turn the contacting flagellar rotor. A derivative model suggests that significant changes in rotor diameter occur during switching, enabling each MotA₅MotB₂ stator unit to shift between internal and external gear configurations, causing clockwise (CW) and counterclockwise (CCW) motor rotation, respectively. However, recent structural work favors a mechanism where the stator units dynamically swing back and forth between the two gear configurations without significant changes in rotor diameter. Given the intricate link between the switching model and the gear mechanism for flagellar rotation, a critical evaluation of the underlying assumptions is crucial for refining switching models. This review scrutinizes key assumptions within prevailing models of flagellar rotation and switching, identifies knowledge gaps, and proposes avenues for future biophysical tests. Full article

A flagellar motor can rotate either counterclockwise (CCW) or clockwise (CW), and rotational switching is triggered by conformational changes in FliG, although the molecular mechanism is still unknown. Here, we found that cheY deletion, which locks motor rotation in the CCW direction, restored the motility abolished by the fliG L259Q mutation. We found that the CCW-biased fliG G214S mutation also restored the swimming of the L259Q mutant, but the CW-biased fliG G215A mutation did not. Since the L259 residue participates in forming the FliG hydrophobic core at its C-terminal domain, mutations were introduced into residues structurally closer to L259, and their motility was examined. Two mutants, D251R and L329Q, exhibited CW-biased rotation. Our results suggest that mutations in the hydrophobic core of FliG_C collapse its conformational switching and/or stator interaction; however, the CCW state of the rotor enables rotation even with this disruption. Full article