TusA is a Versatile Protein That Links Translation Efficiency to Cell Division in Escherichia coli

To enable accurate and efficient translation, sulfur modifications are introduced posttranscriptionally into nucleosides in tRNAs. The biosynthesis of tRNA sulfur modifications involves unique sulfur trafficking systems for the incorporation of sulfur atoms in different nucleosides of tRNA. One of the proteins that is involved in inserting the sulfur for 5-methylaminomethyl-2-thiouridine (mnm5s2U34) modifications in tRNAs is the TusA protein. TusA, however, is a versatile protein that is also involved in numerous other cellular pathways. Despite its role as a sulfur transfer protein for the 2-thiouridine formation in tRNA, a fundamental role of TusA in the general physiology of Escherichia coli has also been discovered. Poor viability, a defect in cell division, and a filamentous cell morphology have been described previously for tusA-deficient cells. In this report, we aimed to dissect the role of TusA for cell viability. We were able to show that the lack of the thiolation status of wobble uridine (U34) nucleotides present on Lys, Gln, or Glu in tRNAs has a major consequence on the translation efficiency of proteins; among the affected targets are the proteins RpoS and Fis. Both proteins are major regulatory factors, and the deregulation of their abundance consequently has a major effect on the cellular regulatory network, with one consequence being a defect in cell division by regulating the FtsZ ring formation. IMPORTANCE More than 100 different modifications are found in RNAs. One of these modifications is the mnm5s2U modification at the wobble position 34 of tRNAs for Lys, Gln, and Glu. The functional significance of U34 modifications is substantial since it restricts the conformational flexibility of the anticodon, thus providing translational fidelity. We show that in an Escherichia coli TusA mutant strain, involved in sulfur transfer for the mnm5s2U34 thio modifications, the translation efficiency of RpoS and Fis, two major cellular regulatory proteins, is altered. Therefore, in addition to the transcriptional regulation and the factors that influence protein stability, tRNA modifications that ensure the translational efficiency provide an additional crucial regulatory factor for protein synthesis.

Role of Cell Surface Hydrophobicity in the Pathogenesis of Medically-Significant Fungi

Cell surface hydrophobicity (CSH) is an important cellular biophysical parameter which affects both cell-cell and cell-surface interactions. In dimorphic fungi, multiple factors including the temperature-induced shift between mold and yeast forms have strong effects on CSH with higher hydrophobicity more common at the lower temperatures conducive to filamentous cell growth. Some strains of Cryptococcus neoformans exhibit high CSH despite the presence of the hydrophilic capsule. Among individual yeast colonies from the same isolate, distinct morphologies can correspond to differences in CSH. These differences in CSH are frequently associated with altered virulence in medically-significant fungi and can impact the efficacy of antifungal therapies. The mechanisms for the maintenance of CSH in pathogenic fungi remain poorly understood, but an appreciation of this fundamental cellular parameter is important for understanding its contributions to such phenomena as biofilm formation and virulence.

Controlled spatial organization of bacterial clusters reveals cell filamentation is vital for Xylella fastidiosa biofilm formation

The morphological plasticity of bacteria to form filamentous cells commonly represents an adaptive strategy induced by stresses. In contrast, for diverse pathogens filamentous cells have been observed during biofilm formation, with function yet to be elucidated. To identify prior hypothesized quorum sensing as trigger of such cell morphogenesis, spatially controlled cell adhesion is pivotal. Here, we demonstrate highly-selective cell adhesion of the biofilm-forming phytopathogen Xylella fastidiosa to gold-patterned SiO2 substrates with well-defined geometries and dimensions. The consequent control of both cell density and distances between cell clusters using these patterns provided evidence of quorum sensing governing filamentous cell formation. While cell morphogenesis is induced by cell cluster density, filamentous cell growth is oriented towards neighboring cell clusters and distance-dependent; large interconnected cell clusters create the early biofilm structural framework. Together, our findings and investigative platform could facilitate therapeutic developments targeting biofilm formation mechanisms of X. fastidiosa and other pathogens.

Vibrio choleraefilamentation promotes chitin surface attachment at the expense of competition in biofilms

Collective behavior in spatially structured groups, or biofilms, is the norm among microbes in their natural environments. Though biofilm formation has been studied for decades, tracing the mechanistic and ecological links between individual cell morphologies and the emergent features of cell groups is still in its infancy. Here we use single-cell–resolution confocal microscopy to explore biofilms of the human pathogenVibrio choleraein conditions mimicking its marine habitat. Prior reports have noted the occurrence of cellular filamentation inV. cholerae, with variable propensity to filament among both toxigenic and nontoxigenic strains. Using a filamenting strain ofV. choleraeO139, we show that cells with this morphotype gain a profound competitive advantage in colonizing and spreading on particles of chitin, the material many marineVibriospecies depend on for growth in seawater. Furthermore, filamentous cells can produce biofilms that are independent of primary secreted components of theV. choleraebiofilm matrix; instead, filamentous biofilm architectural strength appears to derive at least in part from the entangled mesh of cells themselves. The advantage gained by filamentous cells in early chitin colonization and growth is countered in long-term competition experiments with matrix-secretingV. choleraevariants, whose densely packed biofilm structures displace competitors from surfaces. Overall, our results reveal an alternative mode of biofilm architecture that is dependent on filamentous cell morphology and advantageous in environments with rapid chitin particle turnover. This insight provides an environmentally relevant example of how cell morphology can impact bacterial fitness.

Filamentation ofVibrio choleraeis an adaptation for surface attachment and biofilm architecture

Collective behavior in spatially structured groups, or biofilms, is the norm among microbes in their natural environments. Though microbial physiology and biofilm formation have been studied for decades, tracing the mechanistic and ecological links between individual cell properties and the emergent features of cell groups is still in its infancy. Here we use single-cell resolution confocal microscopy to explore biofilm properties of the human pathogenVibrio choleraein conditions closely mimicking its marine habitat. We find that some – but not all – pandemic isolates produce filamentous cells than can be over 50 μm long. This filamentous morphotype gains a profound competitive advantage in colonizing and spreading on particles of chitin, the material many marineVibriospecies depend on for growth outside of hosts. Furthermore, filamentous cells can produce biofilms that are independent of all currently known secreted components of theV. choleraebiofilm matrix; instead, filamentous biofilm architectural strength appears to derive from the entangled mesh of cells themselves. The advantage gained by filamentous cells in early chitin colonization and growth is counter-balanced in longer term competition experiments with matrix-secretingV. choleraevariants, whose densely packed biofilm structures displace competitors from surfaces. Overall our results reveal a novel mode of biofilm architecture that is dependent on filamentous cell morphology and advantageous in environments with rapid chitin particle turnover. This insight provides concrete links betweenV. choleraecell morphology, biofilm formation, marine ecology, and – potentially – the strain composition of cholera epidemics.

Determination of the relationship between epiphytes and selected filamentous bacteria in activated sludge

Activated sludge (AS) flocs are paramount in biological treatment of wastewater, are comprised of microbial consortia with organic and inorganic material bound together by extra polymeric substances (EPS). The filamentous bacteria play a vital role in the floc formation process by providing the necessary structural support. Presence of epiphytic attachment on selected filamentous bacteria is a commonly occurring phenomenon in activated sludge samples. Different theories have been proposed to describe this phenomenon; however, not much research has been carried out to explore the profundity of the attachment. In this study, an attempt has been made to elucidate the intrinsic nature of the epiphytic attachment between the bacterial rods and filamentous bacteria based on microscopic (morphological and structural) analysis. Characterization of these epiphytes were performed using fluorescence in situ hybridization (FISH) at group level using Alpha, Beta and Gamma Proteo-bacterial probes. Morphological characteristics of filament hosts and the bacterial rods at the interface region was assessed using scanning electron microscopy (SEM). The SEM micrographs indicated that the attachment was facilitated by more than the EPS layer. Further ultrastructural examination using transmission electron microscopy (TEM) indicated a possible cell-to-cell interaction between epiphytes and the selected filaments. Fibrillar structures resembling amyloid-like proteins were observed within the filament cell targeted by the epiphytes. An interaction was apparent between the amyloid like proteins and the epiphytes as exhibited by the direction of fibrillar structures pointing towards the approaching epiphytes. Common bacterial appendages such as pili and fimbria were absent at the interface and further noted was the presence of cell membrane extensions on the epiphytic bacteria protruding towards the targeted filamentous cell. The sheath of host filaments however, remained intact and unpenetrated, during colonization. Amyloid-like fibrils at interface may potentially play the role of attachment sites for the attaching epiphytes, as attachment facilitating appendages were not visualized.

Impact of Low Frequency Electromagnetic Field Exposure on the Candida Albicans

Effect of low frequency electromagnetic field on growth of selected microorganism is studied in the article. The diploid fungus that grows both as yeast and filamentous cell was chosen for this research. The theory of ion parametric resonance was taken as the base for studying the influence of electromagnetic field on biological structures. We tested the hypothesis, whether it is possible to observe the change in growth properties of Candida albicans with an AC electromagnetic field tuned to resonance with calcium ions cyclotron frequency.

Designed filamentous cell penetrating peptides: probing supramolecular structure-dependent membrane activity and transfection efficiency

In this work, we will demonstrate the supramolecular assembly of single-chain cationic peptides into stable macromolecular filamentous nanostructures and investigate their supramolecular structure-dependent membrane activity for the development of highly efficient therapeutic carriers.