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Environmental Microbiology Apr 2022Light-induced carotenogenesis in Myxococcus xanthus is controlled by the B -based CarH repressor and photoreceptor, and by a separate intricate pathway involving singlet...
Light-induced carotenogenesis in Myxococcus xanthus is controlled by the B -based CarH repressor and photoreceptor, and by a separate intricate pathway involving singlet oxygen, the B -independent CarH paralogue CarA and various other proteins, some eukaryotic-like. Whether other myxobacteria conserve these pathways and undergo photoregulated carotenogenesis is unknown. Here, comparative analyses across 27 Myxococcales genomes identified carotenogenic genes, albeit arranged differently, with carH often in their genomic vicinity, in all three Myxococcales suborders. However, CarA and its associated factors were found exclusively in suborder Cystobacterineae, with carA-carH invariably in tandem in a syntenic carotenogenic operon, except for Cystobacter/Melittangium, which lack CarA but retain all other factors. We experimentally show B -mediated photoregulated carotenogenesis in representative myxobacteria, and a remarkably plastic CarH operator design and DNA binding across Myxococcales. Unlike the two characterized CarH from other phyla, which are tetrameric, Cystobacter CarH (the first myxobacterial homologue amenable to analysis in vitro) is a dimer that combines direct CarH-like B -based photoregulation with CarA-like DNA binding and inhibition by an antirepressor. This study provides new molecular insights into B -dependent photoreceptors. It further establishes the B -dependent pathway for photoregulated carotenogenesis as broadly prevalent across myxobacteria and its evolution, exclusively in one suborder, into a parallel complex B -independent circuit.
Topics: Bacterial Proteins; DNA; Gene Expression Regulation, Bacterial; Myxococcales; Phosphothreonine; Repressor Proteins
PubMed: 35005822
DOI: 10.1111/1462-2920.15895 -
Philosophical Transactions of the Royal... Mar 2021Social bacteria display complex behaviours whereby thousands of cells collectively and dramatically change their form and function in response to nutrient availability...
Social bacteria display complex behaviours whereby thousands of cells collectively and dramatically change their form and function in response to nutrient availability and changing environmental conditions. In this review, we focus on motility, which supports spectacular transitions based on prey availability across its life cycle. A large body of work suggests that these behaviours require sensory capacity implemented at the single-cell level. Focusing on recent genetic work on a core cellular pathway required for single-cell directional decisions, we argue that signal integration, multi-modal sensing and memory are at the root of decision making leading to multicellular behaviours. Hence, may be a powerful biological system to elucidate how cellular building blocks cooperate to form sensory multicellular assemblages, a possible origin of cognitive mechanisms in biological systems. This article is part of the theme issue 'Basal cognition: conceptual tools and the view from the single cell'.
Topics: Microbial Interactions; Myxococcus xanthus
PubMed: 33487114
DOI: 10.1098/rstb.2019.0755 -
Frontiers in Microbiology 2022The bacterium forms both developmental and vegetative types of biofilms. While the former has been studied on both agar plates and submerged surfaces, the latter has...
The bacterium forms both developmental and vegetative types of biofilms. While the former has been studied on both agar plates and submerged surfaces, the latter has been investigated predominantly on agar surfaces as swarming colonies. Here we describe the development of a microplate-based assay for the submerged biofilms of under vegetative conditions. We examined the impacts of inoculation, aeration, and temperature to optimize the conditions for the assay. Aeration was observed to be critical for the effective development of submerged biofilms by , an obligate aerobic bacterium. In addition, temperature plays an important role in the development of submerged biofilms. It is well established that the formation of submerged biofilms by many bacteria requires both exopolysaccharide (EPS) and the type IV pilus (T4P). EPS constitutes part of the biofilm matrix that maintains and organizes bacterial biofilms while the T4P facilitates surface attachment as adhesins. For validation, we used our biofilm assay to examine a multitude of strains with various EPS and T4P phenotypes. The results indicate that the levels of EPS, but not of piliation, positively correlate with submerged biofilm formation in .
PubMed: 35572678
DOI: 10.3389/fmicb.2022.894562 -
Microorganisms May 2021Myxobacteria are Gram-negative δ-proteobacteria found predominantly in terrestrial habitats and often brightly colored due to the biosynthesis of carotenoids.... (Review)
Review
Myxobacteria are Gram-negative δ-proteobacteria found predominantly in terrestrial habitats and often brightly colored due to the biosynthesis of carotenoids. Carotenoids are lipophilic isoprenoid pigments that protect cells from damage and death by quenching highly reactive and toxic oxidative species, like singlet oxygen, generated upon growth under light. The model myxobacterium turns from yellow in the dark to red upon exposure to light because of the photoinduction of carotenoid biosynthesis. How light is sensed and transduced to bring about regulated carotenogenesis in order to combat photooxidative stress has been extensively investigated in using genetic, biochemical and high-resolution structural methods. These studies have unearthed new paradigms in bacterial light sensing, signal transduction and gene regulation, and have led to the discovery of prototypical members of widely distributed protein families with novel functions. Major advances have been made over the last decade in elucidating the molecular mechanisms underlying the light-dependent signaling and regulation of the transcriptional response leading to carotenogenesis in . This review aims to provide an up-to-date overview of these findings and their significance.
PubMed: 34063365
DOI: 10.3390/microorganisms9051067 -
The Science of the Total Environment Dec 2021The mechanisms of struvite production by biomineralization were investigated for five microorganisms (Bacillus pumilus, Brevibacterium antiquum, Myxococcus xanthus,...
The mechanisms of struvite production by biomineralization were investigated for five microorganisms (Bacillus pumilus, Brevibacterium antiquum, Myxococcus xanthus, Halobacterium salinarum and Idiomarina loihiensis) in municipal wastewater. The microbial exponential phase of growth occurred within the first 48 h of incubation, with growth rates varying from 0.02-0.08 1/h. These five microorganisms removed 23-27 mg/L (66-79%) of ortho-phosphate from wastewater, which was recovered as biological struvite (i.e., bio-struvite) identified by morphological, X-ray diffraction and elemental analysis. Bio-struvite crystals occurred in a low extracellular supersaturation index (0.6-0.8 units). Bio-struvite formation in B. pumilus M. xanthus, H. salinarum cultures was linked to biologically induced mineralization. Whereas B. antiquum and I. loihiensis produced bio-struvite through biologically controlled mineralization mechanism because the crystals presented homogeneity in morphology and size, and intracellular vesicle-like cell structures were observed enclosing electron-dense granules/materials. Nutrient recovery through biomineralization has potential application in wastewater streams promoting circularity within the wastewater industry.
Topics: Alteromonadaceae; Biomineralization; Brevibacterium; Phosphates; Phosphorus; Struvite; Wastewater
PubMed: 34371415
DOI: 10.1016/j.scitotenv.2021.149261 -
MBio Oct 2022In Gram-negative bacteria, secreted polysaccharides have multiple critical functions. In Wzx/Wzy- and ABC transporter-dependent pathways, an outer membrane (OM)...
In Gram-negative bacteria, secreted polysaccharides have multiple critical functions. In Wzx/Wzy- and ABC transporter-dependent pathways, an outer membrane (OM) polysaccharide export (OPX) type translocon exports the polysaccharide across the OM. The paradigm OPX protein Wza of Escherichia coli is an octamer in which the eight C-terminal domains form an α-helical OM pore and the eight copies of the three N-terminal domains (D1 to D3) form a periplasmic cavity. In synthase-dependent pathways, the OM translocon is a 16- to 18-stranded β-barrel protein. In Myxococcus xanthus, the secreted polysaccharide EPS (exopolysaccharide) is synthesized in a Wzx/Wzy-dependent pathway. Here, using experiments, phylogenomics, and computational structural biology, we identify and characterize EpsX as an OM 18-stranded β-barrel protein important for EPS synthesis and identify AlgE, a β-barrel translocon of a synthase-dependent pathway, as its closest structural homolog. We also find that EpsY, the OPX protein of the EPS pathway, consists only of the periplasmic D1 and D2 domains and completely lacks the domain for spanning the OM (herein termed a OPX protein). , EpsX and EpsY mutually stabilize each other and interact in pulldown experiments supporting their direct interaction. Based on these observations, we propose that EpsY and EpsX make up and represent a third type of translocon for polysaccharide export across the OM. Specifically, in this composite translocon, EpsX functions as the OM-spanning β-barrel translocon together with the periplasmic OPX protein EpsY. Based on computational genomics, similar composite systems are widespread in Gram-negative bacteria. Bacteria secrete a wide variety of polysaccharides that have critical functions in, e.g., fitness, surface colonization, and biofilm formation and in beneficial and pathogenic human-, animal-, and plant-microbe interactions. In Gram-negative bacteria, export of these chemically diverse polysaccharides across the outer membrane depends on two known translocons, i.e., an outer membrane OPX protein in Wzx/Wzy- and ABC transporter-dependent pathways and an outer membrane 16- to 18-stranded β-barrel protein in synthase-dependent pathways. Here, using a combination of experiments in Myxococcus xanthus, phylogenomics, and computational structural biology, we provide evidence supporting that a third type of translocon can export polysaccharides across the outer membrane. Specifically, in this translocon, an outer membrane-spanning β-barrel protein functions together with an entirely periplasmic OPX protein that completely lacks the domain for spanning the OM. Computational genomics support that similar composite systems are widespread in Gram-negative bacteria.
Topics: ATP-Binding Cassette Transporters; Bacterial Outer Membrane Proteins; Escherichia coli; Escherichia coli Proteins; Gram-Negative Bacteria; Periplasm; Polysaccharides, Bacterial
PubMed: 35972145
DOI: 10.1128/mbio.02032-22 -
Structure (London, England : 1993) Apr 2022Encapsulins are bacterial organelle-like cages involved in various aspects of metabolism, especially protection from oxidative stress. They can serve as vehicles for a...
Encapsulins are bacterial organelle-like cages involved in various aspects of metabolism, especially protection from oxidative stress. They can serve as vehicles for a wide range of medical applications. Encapsulin shell proteins are structurally similar to HK97 bacteriophage capsid protein and their function depends on the encapsulated cargos. The Myxococcus xanthus encapsulin system comprises EncA and three cargos: EncB, EncC, and EncD. EncB and EncC are similar to bacterial ferritins that can oxidize Fe to less toxic Fe. We analyzed EncA, EncB, and EncC by cryo-EM and X-ray crystallography. Cryo-EM shows that EncA cages can have T = 3 and T = 1 symmetry and that EncA T = 1 has a unique protomer arrangement. Also, we define EncB and EncC binding sites on EncA. X-ray crystallography of EncB and EncC reveals conformational changes at the ferroxidase center and additional metal binding sites, suggesting a mechanism for Fe oxidation and storage within the encapsulin shell.
Topics: Bacterial Proteins; Crystallography, X-Ray; Ferritins; Iron; Myxococcus xanthus
PubMed: 35150605
DOI: 10.1016/j.str.2022.01.008 -
Microorganisms Oct 2021Predator impacts on prey diversity are often studied among higher organisms over short periods, but microbial predator-prey systems allow examination of prey-diversity...
Predator impacts on prey diversity are often studied among higher organisms over short periods, but microbial predator-prey systems allow examination of prey-diversity dynamics over evolutionary timescales. We previously showed that commonly evolved minority mucoid phenotypes in response to predation by the bacterial predator by one time point of a coevolution experiment now named MyxoEE-6. Here we examine mucoid frequencies across several MyxoEE-6 timepoints to discriminate between the hypotheses that mucoids were increasing to fixation, stabilizing around equilibrium frequencies, or heading to loss toward the end of MyxoEE-6. In four focal coevolved prey populations, mucoids rose rapidly early in the experiment and then fluctuated within detectable minority frequency ranges through the end of MyxoEE-6, generating frequency dynamics suggestive of negative frequency-dependent selection. However, a competition experiment between mucoid and non-mucoid clones found a predation-specific advantage of the mucoid clone that was insensitive to frequency over the examined range, leaving the mechanism that maintains minority mucoidy unresolved. The advantage of mucoidy under predation was found to be associated with reduced population size after growth (productivity) in the absence of predators, suggesting a tradeoff between productivity and resistance to predation that we hypothesize may reverse mucoid vs non-mucoid fitness ranks within each MyxoEE-6 cycle. We also found that mucoidy was associated with diverse colony phenotypes and diverse candidate mutations primarily localized in the exopolysaccharide operon . Collectively, our results show that selection from predatory bacteria can generate apparently stable sympatric phenotypic polymorphisms within coevolving prey populations and also allopatric diversity across populations by selecting for diverse mutations and colony phenotypes associated with mucoidy. More broadly, our results suggest that myxobacterial predation increases long-term diversity within natural microbial communities.
PubMed: 34683400
DOI: 10.3390/microorganisms9102079 -
Molecular Microbiology Oct 2021Exopolysaccharide (EPS) layers on the bacterial cell surface are key determinants of biofilm establishment and maintenance, leading to the formation of higher-order 3D...
Exopolysaccharide (EPS) layers on the bacterial cell surface are key determinants of biofilm establishment and maintenance, leading to the formation of higher-order 3D structures that confer numerous survival benefits to a cell community. In addition to a specific cell-associated EPS glycocalyx, we recently revealed that the social δ-proteobacterium Myxococcus xanthus secretes a novel biosurfactant polysaccharide (BPS) to the extracellular milieu. Together, secretion of the two polymers (EPS and BPS) is required for type IV pilus (T4P)-dependent swarm expansion via spatio-specific biofilm expression profiles. Thus the synergy between EPS and BPS secretion somehow modulates the multicellular lifecycle of M. xanthus. Herein, we demonstrate that BPS secretion functionally alters the EPS glycocalyx via destabilization of the latter, fundamentally changing the characteristics of the cell surface. This impacts motility behaviors at the single-cell level and the aggregative capacity of cells in groups via cell-surface EPS fibril formation as well as T4P production, stability, and positioning. These changes modulate the structure of swarm biofilms via cell layering, likely contributing to the formation of internal swarm polysaccharide architecture. Together, these data reveal the manner by which the combined secretion of two distinct polymers induces single-cell changes that modulate swarm biofilm communities.
Topics: Bacterial Proteins; Biofilms; Cell Membrane; Fimbriae, Bacterial; Glycocalyx; Myxococcus xanthus; Polysaccharides, Bacterial
PubMed: 34455651
DOI: 10.1111/mmi.14803 -
RSC Chemical Biology Nov 20238-Azido-3,8-dideoxy-α/β-d--oct-2-ulosonic acid (Kdo-8-N) is a Kdo derivative used in metabolic labeling of lipopolysaccharide (LPS) structures found on the cell...
8-Azido-3,8-dideoxy-α/β-d--oct-2-ulosonic acid (Kdo-8-N) is a Kdo derivative used in metabolic labeling of lipopolysaccharide (LPS) structures found on the cell membrane of Gram-negative bacteria. Several studies have reported successful labeling of LPS using Kdo-8-N and visualization of LPS by a fluorescent reagent through click chemistry on a selection of Gram-negative bacteria such as strains, , and . Motivated by the promise of Kdo-8-N to be useful in the investigation of LPS biosynthesis and cell surface labeling across different strains, we set out to explore the variability in nature and efficiency of LPS labeling using Kdo-8-N in a variety of strains and serotypes. We optimized the chemical synthesis of Kdo-8-N and subsequently used Kdo-8-N to metabolically label pathogenic strains from commercial and clinical origin. Interestingly, different extents of labeling were observed in different strains, which seemed to be dependent also on growth media, and the majority of labeled LPS appears to be of the 'rough' LPS variant, as visualized using SDS-PAGE and fluorescence microscopy. This knowledge is important for future application of Kdo-8-N in the study of LPS biosynthesis and dynamics, especially when working with clinical isolates.
PubMed: 37920390
DOI: 10.1039/d3cb00110e