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Putative evolution of Myxococcus fulvus 124B02 plasmid pMF1 from a chromosomal segment in another Myxococcus species.

Myxobacteria or order Myxococcales (old nomenclature) or phylum Myxococcota (new terminology) are fascinating organisms well known for their diverse peculiar physiological, taxonomic, and genomic properties. Researchers have long sought to identify plasmids within these organisms, yet thus far, only two organisms from different families have been found to harbor a plasmid. This study delves into the putative evolution of one of these plasmids, i.e., pMF1 present in Myxococcus fulvus 124B02 in the suborder Cystobacterineae and family Myxococcaceae. Here, we first reannotated the pMF1 plasmid genome sequence and identified two additional open reading frames or putative genes which were not annotated until now. We further reported that all pMF1 plasmid genes depict homology with Myxococcus stipitatus CYD1 draft genome (contig 28) and a chromosomal segment of M. stipitatus DSM14675 in a syntenic manner, implying the presence of plasmid-like structure in M. stipitatus CYD1, integrated into its chromosome. To comprehend the relationship among these three species, we conducted phylogenetic analyses using 16S and concatenated housekeeping genes and genome-to-genome distance calculator (GGDC) analysis, which confirmed that M. stipitatus CYD1 is a distinct and novel species within the genus Myxococcus. Overall, this comparative genomic study sheds light on the putative emergence of the pMF1 plasmid from a common ancestor of closely related yet distinct species, M. stipitatus CYD1, possibly through the partition from its chromosome as a segment.IMPORTANCEMyxobacteria are not well known to have plasmids. Until now, only two organisms have been shown to have plasmids, raising a pertinent question about how these plasmids evolved randomly within the phylum Myxococcota. The study presented in this manuscript delves into the emergence of the pMF1 plasmid found in Myxococcus fulvus 124B02, a member of the suborder Cystobacterineae and family Myxococcaceae. Our research addresses this intriguing topic of plasmid identification and evolution within myxobacteria, which are a group of fascinating organisms that have garnered significant interest due to their diverse physiological, taxonomic, and genomic properties.

Plasmids

Site of ATPase activity in Myxococcus xanthus: lipid requirement for enzyme activity. Dedicated to Professor Dr. W. Schwartz on his 80th birthday.

Treatment of cells with lysophosphatidylcholine, lysozyme, and phospholipase D removed most of their phospholipids and reduced ATPase activity to near zero. Addition of a microdispersion of phospholipids restored enzyme activity to various degrees. Phosphatidylcholine was most effective in reconstitution experiments, less effective were phosphatidylethanolamine and phosphatidylserine. Lipid analyses of cell fractions were possible through separation of cell wall and cell membrane in a sucrose gradient after differentiated treatment of glutaraldehyde fixed cells with lysophosphatidylcholine, lysozyme, and pronase. Phosphatidylcholine was almost exclusively a component of the cell membrane, whereas phosphatidylethanolamine was that of the wall. It is concluded that lipids are necessary for in vivo function of a Mg-dependent ATPase, and that membrane-associated phosphatidylcholine may serve as a matrix for the enzyme. Lipid extracts made from cells or cell fractions contained plasmologens, not previously reported to occur in Gram-negative, aerobic bacteria.

Adenosine Triphosphatases

Evidence for repair of ultraviolet-induced damage in Cystobacter species (Myxobacterales).

Cystobacter species strain CK 1 does not grow with more than 0.2 microgram/ml acriflavine. Spontaneous two-step mutants growing with 2 microgram acriflavine per ml have been selected. One mutant (strain CK3) was used to investigate the effect of repair inhibitors. Both strains exhibit pronounced shoulders in their UV dose curves of inactivation. Acriflavine (AF), coumarin (CU), and caffeine (CA) when incorporated in the post-irradiation plating medium decreased survival of irradiated cells. Post-treatment with 2 microgram acriflavine/ml abolished the shoulder of the curve. Caffeine (1600 microgram/ml) and coumarin (350 microgram/ml) reduced it only to about 40%. It is concluded that probably two repair mechanisms are present. Pre-treatment of the cells with 2 microgram acriflavine/ml for two hours before UV-irradiation resulted in a constant dose enhancement factor of 1.9. The protective effect is increased with the time of treatment with acriflavine. This may indicate that pyrimidine dimers are responsible for UV-inactivation.

Acriflavine

[Occurrence and distribution of fruiting body-forming myxobacteria in Siebengebirge. Comparative studies with special reference to characteristic biotypes].

In the region of the "Siebengebirge" near Bonn, six characteristic biotopes were studied with respect to the occurrence of fruiting body forming myxobacteria. 23 different species, belonging to the 8 known genera, were found. The number of species in the respective biotopes varied between 6 and 17, the average numbers of species per sampling area were between 2.1 and 10.0. 15 species were found on dung pellets of wild living herbivorous mammals. Deciduous forest soils contained 13 species in spring, 17 species in autumn. Rock coverings contained 14, creek slopes 16, bark of living trees 9, and rotting wood 16 fruiting myxobacterial species. Myxococcus fulvus, Myxococcus coralloides and Archangium gephyra occurred in all biotops studied. The rare species were Melittangium boletus, Polyangium vitellinum, Stigmatella aurantiaca, and Chondramyces apiculatus. The composition of the myxobacterial flora depends on ecological factors (kind of dung pellets, rock, bark and pH). The ecological influences upon the distribution are discussed.

Altitude

Purification and effects of fulvocin C, a bacteriocin from Myxococcus fulvus Mx f16.

Fulvocin C is a bacteriocin from Myxococcus fulvus Mx f16. It has a molecular weight of 4672 and is one of the smallest bacteriocins known. Four disulfide bonds give the molecule a tight structure, so that its native form was not attacked by chymotrypsin or pronase. Fulvocin C was stable in various organic solvents and could tolerate 80 degrees C in aqueous solution without loss of activity. The killing effect of fulvocin C was observed only at concentrations higher than 0.25 mumol/1. Macromolecular synthesis (DNA, RNA, protein) was affected very gradually. Viability in growing cultures decreased slowly from 100 to 25% during one generation (8 h). Cell division was affected early. After one generation v-shaped cell pairs had accumulated in the culture. Electron microscopic pictures revealed extended membrane systems connected with the inner membrane. The most striking effect was that often the outer membranes of neighbouring cells seemed to have fused laterally. With further incubation many cells lost their rod shape and empty bags became predominant.

Bacterial Proteins

Myxobacterial slime and proteolytic activity.

An extracellular protein-polysaccharide-lipide (PPL) complex from exponentially growing cultures of Myxococcus virescens was purified by phosphate precipitation and gel chromatography. The high molecular weight slime polymer appeared homogenous upon isoelectric focusing. The PPL complex exhibited proteolytic activity against gelatin and the activity was only partly reduced by heat treatment. The function of the slime polymer as protein denatured was studied. The complex formed micelles similar to anionic detergents and it inhibited the precipitation and coagulation of proteins by trichloroacetic acid. Lysozyme was totally inactivated when treated with the PPL complex. By gel chromatography binding studies, the PPL complex was found to bind lysozyme in the ratio of 1 to 5.8 (w/w). After separation of added protein from the complex the anticoagulation effect on the protein remained. The biological function of the PPL complex was demonstrated with hemoglobin. When all susceptible peptide bonds in PPL-treated hemoglobin were hydrolyzed by trypsin only 20% in the urea-denatured protein were attacked. The combined role of slime and proteolytic activity is discussed.

Bacterial Proteins

Morphogenesis in Myxococcus xanthus and Myxococcus virescens Myxobacterales.

1. Myxococcus xanthus B and M. virescens V2 were compared with a view to establishing the control of their morphogenetic cycles. Both organisms are typical myxococci and on solid media with low concentrations of nutrient they form fruiting bodies, within which vegetative cells convert to myxospores. Ultrathin sections of vegetative M. virescens resembled those of M. xanthus and contained prominent heavily stained bodies, presumed to be polyphosphate granules. Shadowed preparations showed fimbriae associated with M. xanthus but not with M. virescens. 2. M. xanthus B converted to myxospores in liquid medium in response to certain alcohols. M. virescens V2 produced phase-refractile spheres, which were not viable and had an unusual ultrastructure. 3. The distributions of fruiting bodies on solid media containing 0.02% Casitone were recorded for the two species and were compared with a Poisson distribution. Cells responded to differences in cell density in a manner suggestive of a response to a chemotactic attractant. Cells growing vegetatively and also cells forming fruiting bodies produced 3',5'-cyclic adenosine monophosphate (cAMP) as measured by the incorporation of exogeneous [3H] adenosine into cAMP. 4. The significance of these findings for theories of fruiting body formation are discussed.

Chemotaxis

Bacteriocins from Myxococcus fulvus (Myxobacterales).

Bacteriocin-like activities were found in several Myxococcus fulvus strains. One strain, Mx f16, exerted strong inhibitory effects on several myxobacterial strains. Synthesis of its bacteriocinic activity could not be induced by mitomycin. Electrophoresis and molecular sieve chromatography revealed at least three different bacteriocinic substances of low molecular weight.

Bacteriocins

Studies on the cell cycle of Myxobacter AL-1. II. Activities of seven enzymes during the cell cycle.

The properties of seven enzymes were studied in extracts from Myxobacter AL-1. The enzymes were isocitrate dehydrogenase (E.C.1.1.1.42), succinate dehydrogenase (E.C.1.3.99.1), alkaline phosphatase (E.C.3.1.3.1), alpha-glucosidase (E.C.3.2.1.20), beta-glucosidase (E.C.3.2.1.21), beta-galactosidase (E.C.3.2.1.23), and N-acetyl-glucosaminidase (E.C. 3.2.1.30). Four of these enzymes: isocitrate dehydrogenase, alpha-glucosidase, beta-glucosidase, and beta-galactosidase are cytosolic enzymes. Succinate dehydrogenase was found to be located on the cytoplasmic membrane system, whereas alkaline phosphatase and N-acetylglucosaminidase were considered as enzymes which bind the outer membranes resp. the cell wall. During the cell cycle, all enzymes have a pattern of discontinuous activity increase. Succinate dehydrogenase and isocitrate dehydrogenase exhibit a stepwise increase of activity, whereas the other enzymes follow the pattern of a peak enzyme.

Acetylglucosaminidase

On the structure of the peptidoglycan of cell walls from Myxobacter AL-1 (myxobacterales).

Basically the peptidoglycan of Myxobater AL-1 consists of alternating beta-1,4-linked N-acetylglucosamic-N-acetylmuramic acid chains. After splitting the aminosugar backbone with a specific algal enzyme three subunits arise: a monomer, a dimer and a timer. Investigation of the monomer with specific enzymes and comparison of the degradation products to standards derived from other bacterial peptidoglycans suggest the following structure of the monomer peptide: L-alanyl-D-glutamic-L-meso-diaminopimelic-D-alanine. A D-alanyl-D-meso-diaminopimelic acid bond is the bridgebond between the peptides of the subunits.

Alanine

Mutants of Myxococcus xanthus insensitive to glycerol-induced myxospore formation.

Mutants of Myxococcus xanthus FBt unable to form myxospores in response to 0.5 M glycerol arise spontaneously with a frequency of 1--3 X 10(-5). These mutants are designated glc. Ultraviolet mutagenesis increases the frequency to a maximum of 7% of the survivors. The reversion frequency following ultraviolet irradiation of spontaneous glc mutants is less than 10(-3). Of four glc mutants examined, none form myxospores in response to the alternative inducers, ethylene glycol and dimethyl sulphoxide. One glc mutant is induced by 1.5 M glycerol; strain FBt responds to this glycerol concentration with low efficiency myxospore formation. Strain FBt and glc mutants all produce myxospores with low efficiency in response to phenyl ethanol. Of 117 glc mutants tested, 109 form fruiting bodies containing mature myxospores; thus, mutations to the glc phenotype do not normally block myxospore formation within the fruiting cycle of the organism.

Dimethyl Sulfoxide

Deoxyribonucleic acid base composition of Simonsiellaceae.

The molar percentages of guanine plus cytosine in the DNA of 51 strains of Simonsiellaceae were determined by buoyant density ultracentrifugation of cell lysates in CsCl. The DNA base ratios ranged from 41-55 mole-% guanine plus cytosine. These values fall within the range known for the Order Cytophagales, the non-fruiting gliding bacteria, and are outside the range of the Order Myxobacterales, the fruiting myxobacteria. Among the strains of the genus Simonsiella, four distinct group can be delineated on the basis of source of origin (sheep, dog, cat, human) and GC content. The neotype of Alysiella filiformis has a GC content of 45.4 mole-%.

Animals

South African Myxococcota: an untapped resource for microbial ecolo gy and biotechnology.

An extraordinary multicellular life cycle, ecological versatility, and prolific production of bioactive secondary metabolites characterise the phylum Myxococcota. While research has predominantly focused on Myxococcota in Asia, Europe, and North America, their potential occurrence in Sub-Saharan Africa remains largely unexplored. To date, only one study has isolated Myxococcota in South Africa, with additional findings limited to incidental detection through metagenomic studies. Considering South Africa's ecological diversity, its biomes may represent promising but under-examined environments for systematic bioprospecting aimed at discovering novel Myxococcota with ecological or biotechnological potential. The recent reclassification of Myxococcota from the former Deltaproteobacteria has provided a more coherent taxonomic framework to guide future ecological and systematic studies. This review presents an overview of the taxonomic revision and explores the potential occurrence of Myxococcota in South African biomes. It covers the challenges associated with conventional culture-based isolation methods and highlights potential genome- and metagenome-based approaches, including the use of metagenome-assembled genomes (MAGs) to identify cryptic biosynthetic gene clusters (BGCs), while acknowledging current limitations. Considering the increasing resistance to chemical fungicides in South African agriculture, this review further explores the potential of Myxococcota-derived secondary metabolites as candidate bioprotective alternatives. By identifying current research gaps, it aims to support future efforts towards systematic bioprospecting to investigate the ecological and biotechnological potential of Myxococcota in South Africa. KEY POINTS: • South African biomes may harbour novel Myxococcota with biosynthetic potential. • Genome mining could reveal cryptic biosynthetic gene clusters (BGCs). • Myxococcota metabolites may help control resistant fungal phytopathogens.

South Africa

Aspartokinase isoenzymes of the fruiting myxobacterium Myxoccus xanthus.

Two isoenzymes of aspartokinase can be found in extracts of the differentiating bacterium Myxococcus xanthus. Aspartokinase I is repressed by L-lysine and feedback is inhibited by meso-diaminopimelate and by low concentrations of L-lysine. However, the inhibition by L-lysine is no longer observed at high concentration of this amino acid. Aspartokinase II is repressed and feedback inhibited specifically by L-threonine. Both enzymes are stimulated significantly by L-methionine and L-isoleucine; the effect is greater with aspartokinase I. The role of these enzymes in relation to growth conditions of the organism is discussed and a correlation with life cycle activity is indicated.

Aspartate Kinase