scholarly journals Multicellular String-Like Structure Formation by Salmonella Typhimurium Depends on Cellulose Production: Roles of Diguanylate Cyclases, YedQ and YfiN

2020 ◽  
Vol 11 ◽  
Author(s):  
Alan Varghese ◽  
Semanti Ray ◽  
Taru Verma ◽  
Dipankar Nandi

Bacteria face diverse stresses in the environment and, sometimes, respond by forming multi-cellular structures, e.g., biofilms. Here, we report a novel macroscopic and multi-cellular structure formed by Salmonella Typhimurium, which resembles small strings. These string-like structures, ∼1 cm long, are induced under some stress conditions: iron deprivation by 2,2-Bipyridyl or low amounts of antibiotics or ethanol in minimal media. However, cells in strings revert back to planktonic growth upon return to nutrient rich media. Compared to planktonic cells, strings are more resistant to antibiotics and oxidative stress. Also, strains lacking csgD or rpoS, which are defective in the classical rdar biofilm formation, form strings. Furthermore, some biofilm inducing conditions do not result in strings and vice-versa, demonstrating that strings are not related to classical CsgD-dependent biofilms. Cells in a string are held together by cellulose and a strain lacking bcsA, which is defective in cellulose production, does not form strings. In addition, reductive stress conditions such as dithiothreitol (DTT) or mutations in the Disulfide bonding system (DSB) also give rise to strings. The amounts of c-di-GMP are increased upon string formation and studies with single and double deletion strains of the diguanylate cyclases, yedQ (STM1987) primarily and yfiN (STM2672) partly, revealed their importance for string formation. This is the first study showcasing the ability of Salmonella to produce high amounts of cellulose in liquid culture, instead of an interface, in a CsgD-independent manner. The relevance and possible applications of strings in the production of bacterial cellulose and bioremediation are discussed.

2014 ◽  
Vol 196 (21) ◽  
pp. 3690-3699 ◽  
Author(s):  
D. A. Hufnagel ◽  
W. H. DePas ◽  
M. R. Chapman

mBio ◽  
2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Sandra Kunz ◽  
Anke Tribensky ◽  
Wieland Steinchen ◽  
Luis Oviedo-Bocanegra ◽  
Patricia Bedrunka ◽  
...  

ABSTRACT Bacillus subtilis contains two known cyclic di-GMP (c-di-GMP)-dependent receptors, YdaK and DgrA, as well as three diguanylate cyclases (DGCs): soluble DgcP and membrane-integral DgcK and DgcW. DgrA regulates motility, while YdaK is responsible for the formation of a putative exopolysaccharide, dependent on the activity of DgcK. Using single-molecule tracking, we show that a majority of DgcK molecules are statically positioned in the cell membrane but significantly less so in the absence of YdaK but more so upon overproduction of YdaK. The soluble domains of DgcK and of YdaK show a direct interaction in vitro, which depends on an intact I-site within the degenerated GGDEF domain of YdaK. These experiments suggest a direct handover of a second messenger at a single subcellular site. Interestingly, all three DGC proteins contribute toward downregulation of motility via the PilZ protein DgrA. Deletion of dgrA also affects the mobility of DgcK within the membrane and also that of DgcP, which arrests less often at the membrane in the absence of DgrA. Both, DgcK and DgcP interact with DgrA in vitro, showing that divergent as well as convergent direct connections exist between cyclases and their effector proteins. Automated determination of molecule numbers in live cells revealed that DgcK and DgcP are present at very low copy numbers of 6 or 25 per cell, respectively, such that for DgcK, a part of the cell population does not contain any DgcK molecule, rendering signaling via c-di-GMP extremely efficient. IMPORTANCE Second messengers are free to diffuse through the cells and to activate all responsive elements. Cyclic di-GMP (c-di-GMP) signaling plays an important role in the determination of the life style transition between motility and sessility/biofilm formation but involves numerous distinct synthetases (diguanylate cyclases [DGCs]) or receptor pathways that appear to act in an independent manner. Using Bacillus subtilis as a model organism, we show that for two c-di-GMP pathways, DGCs and receptor molecules operate via direct interactions, where a synthesized dinucleotide appears to be directly used for the protein-protein interaction. We show that very few DGC molecules exist within cells; in the case of exopolysaccharide (EPS) formation via membrane protein DgcK, the DGC molecules act at a single site, setting up a single signaling pool within the cell membrane. Using single-molecule tracking, we show that the soluble DGC DgcP arrests at the cell membrane, interacting with its receptor, DgrA, which slows down motility. DgrA also directly binds to DgcK, showing that divergent as well as convergent modules exist in B. subtilis. Thus, local-pool signal transduction operates extremely efficiently and specifically.


2015 ◽  
Vol 82 (4) ◽  
pp. 1237-1248 ◽  
Author(s):  
Kimberly N. Cowles ◽  
David K. Willis ◽  
Tyler N. Engel ◽  
Jeffrey B. Jones ◽  
Jeri D. Barak

ABSTRACTIncreasing evidence indicates that despite exposure to harsh environmental stresses,Salmonella entericasuccessfully persists on plants, utilizing fresh produce as a vector to animal hosts. Among the importantS. entericaplant colonization factors are those involved in biofilm formation.S. entericabiofilm formation is controlled by the signaling molecule cyclic di-GMP and represents a sessile lifestyle on surfaces that protects the bacterium from environmental factors. Thus, the transition from a motile, planktonic lifestyle to a sessile lifestyle may represent a vital step in bacterial success. This study examined the mechanisms ofS. entericaplant colonization, including the role of diguanylate cyclases (DGCs) and phosphodiesterases (PDEs), the enzymes involved in cyclic di-GMP metabolism. We found that two biofilm components, cellulose and curli, are differentially required at distinct stages in root colonization and that the DGC STM1987 regulates cellulose production in this environment independent of AdrA, the DGC that controls the majority ofin vitrocellulose production. In addition, we identified a new function for AdrA in the transcriptional regulation of colanic acid and demonstrated thatadrAand colanic acid biosynthesis are associated withS. entericadesiccation tolerance on the leaf surface. Finally, two PDEs with known roles in motility, STM1344 and STM1697, had competitive defects in the phyllosphere, suggesting that regulation of motility is crucial forS. entericasurvival in this niche. Our results indicate that specific conditions influence the contribution of individual DGCs and PDEs to bacterial success, perhaps reflective of differential responses to environmental stimuli.


Author(s):  
Baodi Dai ◽  
Yinxing Xu ◽  
Hongyu Wu ◽  
Jiangye Chen

Abstract Candida albicans has long been known to switch between white and opaque phases; however, a third cell type, referred to as the ‘gray’ phenotype, was recently characterized. The three phenotypes have different colonial morphologies, with white cells forming white-colored colonies and opaque and gray cells forming dark-colored colonies. We previously showed that Wor1-upregulated ferroxidases (Fets) function as pigment multicopper oxidases that regulate the production of dark-pigmented melanin in opaque cells. In this study, we demonstrated that Fets also contributed to dark pigment formation in gray colonies but in a Wor1-independent manner. Deletion of both WOR1 and EFG1 locked cells in the gray phenotype in some rich media. However, the efg1/efg1 wor1/wor1 mutant could switch between white and gray in minimal media depending on the ambient pH. Specifically, mutant cells exhibited the white phenotype at pH 4.5 but switched to gray at pH 7.5. Consistent with phenotype switching, Fets expressions and melanin production were also regulated by ambient pH. Ectopic expression of the Rim101-405 allele in the mutant enabled the pH restriction to be bypassed and promoted gray cell formation in acidic media. Our data suggest that Rim101-upregulated Fets contribute to dark pigment formation in the gray cells.


Crisis ◽  
2012 ◽  
Vol 33 (2) ◽  
pp. 106-112 ◽  
Author(s):  
Christopher M. Bloom ◽  
Shareen Holly ◽  
Adam M. P. Miller

Background: Historically, the field of self-injury has distinguished between the behaviors exhibited among individuals with a developmental disability (self-injurious behaviors; SIB) and those present within a normative population (nonsuicidal self-injury; NSSI),which typically result as a response to perceived stress. More recently, however, conclusions about NSSI have been drawn from lines of animal research aimed at examining the neurobiological mechanisms of SIB. Despite some functional similarity between SIB and NSSI, no empirical investigation has provided precedent for the application of SIB-targeted animal research as justification for pharmacological interventions in populations demonstrating NSSI. Aims: The present study examined this question directly, by simulating an animal model of SIB in rodents injected with pemoline and systematically manipulating stress conditions in order to monitor rates of self-injury. Methods: Sham controls and experimental animals injected with pemoline (200 mg/kg) were assigned to either a low stress (discriminated positive reinforcement) or high stress (discriminated avoidance) group and compared on the dependent measures of self-inflicted injury prevalence and severity. Results: The manipulation of stress conditions did not impact the rate of self-injury demonstrated by the rats. The results do not support a model of stress-induced SIB in rodents. Conclusions: Current findings provide evidence for caution in the development of pharmacotherapies of NSSI in human populations based on CNS stimulant models. Theoretical implications are discussed with respect to antecedent factors such as preinjury arousal level and environmental stress.


Planta Medica ◽  
2014 ◽  
Vol 80 (10) ◽  
Author(s):  
F Nabbie ◽  
O Shperdheja ◽  
J Millot ◽  
J Lindberg ◽  
B Peethambaran

2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Nur Hidaayah

Stress conditions in the elderly means an imbalance condition of biological, psychological, and social are closely related to the response to the threats and dangers faced by the elderly. Pressure or interference that is not fun is usually created when the elderly see a mismatch between the state and the 3 systems available resources. Maintenance actions that need to be done there are 2 types, namely : prevention of exposure to a stressor (precipitation factor) and serious treatment of the imbalance condition/ illness (precipitation factor). Prevention includes: sports, hobbies, friendship, avoid eating foods high in free radicals and harmful substances, sex and setting arrangements adequate rest. Habits of the above if done at a young age to avoid exposure to stress in the elderly. Treatment of the imbalance condition / illness, include : drinking water, meditation, eating fresh fruit, and adequate rest.


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