ABSTRACT
Data on the participation of microbiota in the development of Parkinson's disease allow us to discuss the ability of bacterial preparations to influence the processes leading to neurodegeneration. We studied the effect of oral administration of Limosilactobacillus fermentum U-21 lyophilisate on a model of Parkinson's disease in rats induced by combined intranigral injection of LPS and systemic administration of paraquat. The toxins significantly increased the number of missteps in the "narrowing beam walking" test, but a tendency to a decrease in this parameter was shown after treatment with U-21. It should be noted that U-21 did not reduce the neuronal death in the substantia nigra, but mitigated the inflammatory glial response, decreased the accumulation of phosphorylated α-synuclein and complement protein C3. Our study demonstrated the efficiency of the combined model of parkinsonism and reduction of proinflammatory changes under the influence of pharmabiotic without changes in the nigral neuronal death and motor deficits.
Subject(s)
Disease Models, Animal , Lipopolysaccharides , Substantia Nigra , alpha-Synuclein , Animals , Rats , alpha-Synuclein/metabolism , Male , Substantia Nigra/drug effects , Substantia Nigra/metabolism , Substantia Nigra/pathology , Lipopolysaccharides/pharmacology , Lipopolysaccharides/toxicity , Paraquat , Parkinson Disease/pathology , Parkinson Disease/drug therapy , Parkinson Disease/metabolism , Rats, Wistar , Complement C3/metabolism , Neuroinflammatory Diseases/pathology , Neuroinflammatory Diseases/drug therapyABSTRACT
The global probiotics industry has been undergoing major changes in recent years. Approaches to finding and creating new probiotics, as well as a paradigm of their use in food, medicine, and pharmacology are changing. The catalyst proved to be the increasing popularity and availability of omics technologies, in particular, metagenomic studies of human and animal microbiomes. However, the efficiency and safety of drugs based on probiotic strains, as well as their marketing rates, largely depend on the levels of legal and technical regulation in the field. The present review discusses the aspects of legal regulation in Russia, the European Union and the United States, along with the advantages and disadvantages of probiotics and postbiotics. A consensus is emerging that postbiotics have a number of advantages over classical live probiotic cultures. The review also focuses on the lactobacilli family, which includes the largest number of probiotic strains studied so far and still holds a leading position among probiotics. On the legislative front, Russia is often ahead of its time with adopting such laws as the Federal Law No. 492-FZ on biosecurity, which defined the concept of human and animal microbiota and set forth legislative guidelines for its preservation. The new field of research referred to as microbiome nutrigenomics aims to achieve this goal.
ABSTRACT
OBJECTIVE: To investigate the effects of diet on the gut microbiota and to assess the relationship of these factors with depression. MATERIAL AND METHODS: Microorganisms that predominate in depressed patients were identified and associations of the identified organisms with the patients' diet were performed. Fourteen depressed patients and 14 healthy volunteers with the same socio-demographic parameters were included in the study. The Hamilton Depression Scale, Generalized Anxiety Disorder Questionnaire, and the Center for Epidemiologic Studies Questionnaire were used. RESULTS: Erysipelatoclostridium and Clostridium innocuum species were 11.3 and 14.4 times higher in depressed patients compared with healthy controls. Fusicatenibacter saccharivorans, Faecalibacterium prausnitzii and Roseburia faecis species, as well as members of the genus Roseburia were statistically significantly more abundant in the healthy volunteers group (6.5, 2.14, 8.75 and 5.2 times more frequently compared to patients). The presence of these microorganisms was correlated with dietary components. CONCLUSION: Our study revealed groups of microorganisms that differ in healthy volunteers and depressed patients. The association of these microorganisms with the diet was shown, which partially confirmed the influence of a «healthy diet¼ on the development of depressive disorders.
Subject(s)
Gastrointestinal Microbiome , Depression , Diet , Feces/microbiology , Humans , RNA, Ribosomal, 16SABSTRACT
Today, a number of studies conclusively show that certain bacterial strains, mainly from the genera Lactobacillus and Bifidobacterium, influence the functioning of the central nervous system, leading to changes in beahvior, nociception and the cognitive abilities of humans and animals. Such strains serve as the basis for developing probiotics with a curative potential for the central nervous system - psychobioitcs. However, the question of how to find such strains and which criteria to use for their selection remains unanswered. Some compounds produced by bacteria, such as gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter of the central nervous system, are potential mediators between bacterial cells and the host. Previously, we established that some species of Lactobacillus and Bifidobacterium are capable of producing GABA. We presumed that GABA-producing Lactobacillus and Bifidobacterium strains are great candidates to use as psychobiotics. Therefore, we selected the strains Lactobacillus plantarum 90sk and Bifidobacterium adolescentis 150 as efficient GABA producers. The goal of this work was to assess the probiotic properties of the selected strains as well as their antidepressive effects in mice. We established that the ingestion of the probiotic composition based on the selected strains by BALB/c mice for 2 weeks reduced depressive-like behavior in the forced swimming test; the effect was similar to that of fluoxetine.
Subject(s)
Antidepressive Agents/administration & dosage , Bifidobacterium adolescentis/metabolism , Lactobacillus plantarum/metabolism , Probiotics/administration & dosage , gamma-Aminobutyric Acid/biosynthesis , Animals , Male , Mice , Mice, Inbred BALB CABSTRACT
The article investigates SNP in genes of toxin-antitoxin systems type II in Mycobacterium tuberculosis Beijing lineage strains and their possible role in the development and formation of new sublineages. We established the catalog of SNPs in 142â¯TA systems genes in 1349 sequenced genomes of the M. tuberculosis Beijing lineage. Based on the catalog, 15 new sublineages were identified as part of Beijing lineages by non-synonymous SNP in 21 genes of TA systems. We discovered three toxin genes with mutations specific for epidemiologically dangerous sublineages Beijing-modern (vapC37 A46G, vapC38 T143C) and Beijing-B0/W148 (vapC12 A95G). We proved the functional significance of these polymorphisms by cloning these genes wild-type and with marker mutations for the Beijing lineage vapC12 (A95G), vapC37 (A46G), vapC38 (T143C). In vitro study of their activities revealed effect of mutations on the RNase activity of toxin proteins. Mutations in vapC37 and vapC38 decreased toxin activity, and mutation in the vapC12 increased it. We cloned the toxin vapC37 gene of Mycobacterium smegmatis mc2 155 in both allelic variants: without mutation and with A46G mutation, specific for the Beijing-modern lineage. It was shown that this mutation leads to a loss of toxicity.
Subject(s)
Bacterial Proteins/genetics , Evolution, Molecular , Mycobacterium tuberculosis/genetics , Polymorphism, Single Nucleotide , Toxin-Antitoxin Systems/genetics , Bacterial Proteins/metabolism , Genotype , Mutation , Mycobacterium tuberculosis/growth & development , Mycobacterium tuberculosis/metabolism , Mycobacterium tuberculosis/pathogenicity , Phenotype , Phylogeny , Protein Interaction MapsABSTRACT
The objective of this study was to determine for phosphorylated substrates of the species-specific serine-threonine protein kinase (STPK) Pkb2 from Bifidobacterium longum subsp. longum GT15. Two approaches were employed: analyses of phosphorylated membrane vesicles protein spectra following kinase reactions and analyses of the genes surrounding pkb2. A bioinformatics analysis of the genes surrounding pkb2 found a species-specific gene cluster PFNA in the genomes of 34 different bifidobacterial species. The identified cluster consisted of 5-8 genes depending on the species. The first five genes are characteristic for all considered species. These are the following genes encoding serine-threonine protein kinase (pkb2), fibronectin type III domain-containing protein (fn3), AAA-ATPase (aaa-atp), hypothetical protein with DUF58 domain (duf58) and transglutaminase (tgm). The sixth (protein phosphatase, prpC), seventh (hypothetical protein, BLGT_RS02790), and eighth (FHA domain-containing protein, fha) genes are included in this cluster, but they are not found in all species. The operon organization of the PFNA gene cluster was confirmed with transcriptional analysis. AAA-ATPase, which is encoded by a gene of the PFNA gene cluster, was found to be a substrate of the STPK Pkb2. Fourteen AAA-ATPase sites (seven serine, six threonine, and one tyrosine) phosphorylated by STPK Pkb2 were revealed. Analysis of the spectra of phosphorylated membrane vesicles proteins allowed us to identify eleven proteins that were considered as possible Pkb2 substrates. They belong to several functional classes: proteins involved in transcription and translation; proteins of the F1-domain of the FoF1-ATPase; ABC-transporters; molecular chaperone GroEL; and glutamine synthase, GlnA1. All identified proteins were considered moonlighting proteins. Three out of 11 proteins (glutamine synthetase GlnA1 and FoF1-ATPase alpha and beta subunits) were selected for further in vitro phosphorylation assays and were shown to be phosphorylated by Pkb2. Four phosphorylated substrates of the species-specific STPK Pkb2 from B. longum subsp. longum GT15 were identified for the first time. They included the moonlighting protein glutamine synthase GlnA, FoF1-ATPase alpha and beta subunits, and the chaperone MoxR family of AAA-ATPase. The ability of bifidobacterial STPK to phosphorylate the substrate on serine, threonine, and tyrosine residues was shown for the first time.
Subject(s)
Bifidobacterium longum/enzymology , Bifidobacterium longum/genetics , Multigene Family , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , Computational Biology , Gene Expression Profiling , Operon , Substrate SpecificityABSTRACT
The diversity of Lb. rhamnosus and Lb. fermentum strains isolated from feces, saliva, and the vaginal cavity of 18-22-year-old healthy women residing in central regions of the Russian Federation has been characterized. The results obtained using multilocus sequence typing were identical to those obtained with the analysis of genetic and genomic polymorphism in TA systems. Different as well as identical Lb. rhamnosus and Lb. fermentum sequence types (ST) were isolated from various parts of the body of the same person. Identical ST were also isolated from different women, suggesting that such strains belong to a common pool of strains circulating among the population members. Our results demonstrate that TAs are suitable for characterizing intra-specific diversity of Lb. rhamnosus and Lb. fermentum strains. The advantage of using polymorphisms in TA systems for genotyping is based on the weak number of genes used, and consequently, less time is required for the analysis.
Subject(s)
Antitoxins/genetics , Bacterial Toxins/genetics , Feces/microbiology , Lacticaseibacillus rhamnosus/genetics , Limosilactobacillus fermentum/genetics , Saliva/microbiology , Vagina/microbiology , Adolescent , Adult , Female , Genotype , Humans , Limosilactobacillus fermentum/classification , Limosilactobacillus fermentum/isolation & purification , Lacticaseibacillus rhamnosus/classification , Lacticaseibacillus rhamnosus/isolation & purification , Multilocus Sequence Typing , Polymorphism, Genetic/genetics , Russia , Young AdultABSTRACT
Recent results related to investigation of the role of intestinal microbiota (IM) in development and functioning of the human nervous system are discussed. The role of the microbiota in bidirectional communication between the gastrointestinal tract and the central nervous system is considered. Special attention is paid to the primary IM of infants, which is actively involved in formation of immune and other physiological mechanisms, including the nervous system, and is responsible for the subsequent general and psychical health of a human. The results of research on ability of the commensal intestinal microflora to produce neuroactive compounds, including neurotransmitters, short- and long-chain fatty acids, γ-aminobutyric acid, etc., are summarized. These compounds may have a considerable effect on development and functioning of the central nervous system, including the brain. Research on various animal models is discussed, including investigation of IM effect on behavior, learning abilities and memory, anxiety and depression levels, reaction to emotional stimuli, and stress resistance. A special section deals with probiotic bacteria, which are presently considered as psychobiotics with preventive and therapeutic potential for treatment of neurological and neurophysiological disorders. Development of new paradigms and concepts, rejection of some classical concepts of neurobiology is presently the key condition for the future breakthrough in investigation of human nervous activity.
Subject(s)
Central Nervous System , Gastrointestinal Microbiome/immunology , Neurotransmitter Agents , Central Nervous System/growth & development , Central Nervous System/immunology , Central Nervous System/pathology , Central Nervous System/physiopathology , Humans , Nervous System Diseases/immunology , Nervous System Diseases/microbiology , Nervous System Diseases/physiopathology , Nervous System Diseases/prevention & control , Neurotransmitter Agents/biosynthesis , Neurotransmitter Agents/immunology , Probiotics/pharmacologyABSTRACT
We report a draft genome sequence of Mycobacterium tuberculosis strain B9741 belonging to Beijing B0/W lineage isolated from a HIV patient from Siberia, Russia. This clinical isolate showed MDR phenotype and resistance to isoniazid, rifampin, streptomycin and pyrazinamide. We analyzed SNPs associated with virulence and resistance. The draft genome sequence and annotation have been deposited at GenBank under the accession NZ_LVJJ00000000.
ABSTRACT
Gamma-amino butyric acid (GABA) is an active biogenic substance synthesized in plants, fungi, vertebrate animals and bacteria. Lactic acid bacteria are considered the main producers of GABA among bacteria. GABA-producing lactobacilli are isolated from food products such as cheese, yogurt, sourdough, etc. and are the source of bioactive properties assigned to those foods. The ability of human-derived lactobacilli and bifidobacteria to synthesize GABA remains poorly characterized. In this paper, we screened our collection of 135 human-derived Lactobacillus and Bifidobacterium strains for their ability to produce GABA from its precursor monosodium glutamate. Fifty eight strains were able to produce GABA. The most efficient GABA-producers were Bifidobacterium strains (up to 6 g/L). Time profiles of cell growth and GABA production as well as the influence of pyridoxal phosphate on GABA production were studied for L. plantarum 90sk, L. brevis 15f, B. adolescentis 150 and B. angulatum GT102. DNA of these strains was sequenced; the gadB and gadC genes were identified. The presence of these genes was analyzed in 14 metagenomes of healthy individuals. The genes were found in the following genera of bacteria: Bacteroidetes (Bacteroides, Parabacteroides, Alistipes, Odoribacter, Prevotella), Proteobacterium (Esherichia), Firmicutes (Enterococcus), Actinobacteria (Bifidobacterium). These data indicate that gad genes as well as the ability to produce GABA are widely distributed among lactobacilli and bifidobacteria (mainly in L. plantarum, L. brevis, B. adolescentis, B. angulatum, B. dentium) and other gut-derived bacterial species. Perhaps, GABA is involved in the interaction of gut microbiota with the macroorganism and the ability to synthesize GABA may be an important feature in the selection of bacterial strains - psychobiotics.
Subject(s)
Bacterial Proteins/genetics , Bifidobacterium/genetics , Gastrointestinal Microbiome/genetics , Glutamate Decarboxylase/genetics , Lactobacillus/genetics , Membrane Proteins/genetics , gamma-Aminobutyric Acid/biosynthesis , Bacterial Proteins/metabolism , Bacteroidetes/drug effects , Bacteroidetes/genetics , Bacteroidetes/isolation & purification , Bacteroidetes/metabolism , Bifidobacterium/drug effects , Bifidobacterium/isolation & purification , Bifidobacterium/metabolism , DNA, Bacterial/genetics , Firmicutes/drug effects , Firmicutes/genetics , Firmicutes/isolation & purification , Firmicutes/metabolism , Gastrointestinal Microbiome/drug effects , Gastrointestinal Tract/microbiology , Gene Expression , Glutamate Decarboxylase/metabolism , Humans , Lactobacillus/drug effects , Lactobacillus/isolation & purification , Lactobacillus/metabolism , Membrane Proteins/metabolism , Metagenome , Proteobacteria/drug effects , Proteobacteria/genetics , Proteobacteria/isolation & purification , Proteobacteria/metabolism , Pyridoxal Phosphate/metabolism , Pyridoxal Phosphate/pharmacology , Sodium Glutamate/metabolism , Sodium Glutamate/pharmacologyABSTRACT
The paper provides the annotation and data on sequencing the antibiotic resistance genes in Streptomyces fradiae strain ATCC19609, highly sensitive to different antibiotics. Genome analysis revealed four groups of genes that determined the resistome of the tested strain. These included classical antibiotic resistance genes (nine aminoglycoside phosphotransferase genes, two beta-lactamase genes, and the genes of puromycin N-acetyltransferase, phosphinothricin N-acetyltransferase, and aminoglycoside acetyltransferase); the genes of ATP-dependent ABC transporters, involved in the efflux of antibiotics from the cell (MacB-2, BcrA, two-subunit MDR1); the genes of positive and negative regulation of transcription (whiB and padR families); and the genes of post-translational modification (serine-threonine protein kinases). A comparative characteristic of aminoglycoside phosphotransferase genes in S. fradiae ATCC19609, S. lividans TK24, and S. albus J1074, the causative agent of actinomycosis, is provided. The possibility of using the S. fradiae strain ATCC19609 as the test system for selection of the macrolide antibiotic oligomycin A derivatives with different levels of activity is demonstrated. Analysis of more than 20 semisynthetic oligomycin A derivatives made it possible to divide them into three groups according to the level of activity: inactive (>1 nmol/disk), 10 substances; with medium activity level (0.051 nmol/disk), 12 substances; and more active (0.010.05 nmol/disk), 2 substances. Important for the activity of semisynthetic derivatives is the change in the position of the 33rd carbon atom in the oligomycin A molecule.
Subject(s)
Bacterial Proteins , Drug Resistance, Bacterial , Genome, Bacterial , Oligomycins/pharmacology , Streptomyces , Whole Genome Sequencing , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Bacterial/drug effects , Drug Resistance, Bacterial/genetics , Microbial Sensitivity Tests , Streptomyces/genetics , Streptomyces/metabolismABSTRACT
Previously, we identified six serine/threonine protein kinases (STPK) of Bifidobacterium and named them Pkb1-Pkb6. In the present study, we optimized methods for isolation of the six STPK catalytic domains proteins of B. longum B379M: a method for isolation of Pkb3 and Pkb4 in native conditions, a method for isolation of Pkb5 in denaturing conditions, and a method for isolation of Pkb1, Pkb2, and Pkb6 from inclusion bodies. The dialysis conditions for the renaturation of the proteins were optimized. All of the enzymes were isolated in quantities sufficient for study of the protein activity. The proteins were homogeneous according to SDS-PAGE. The autophosphorylation ability of Pkb1, Pkb3, Pkb4, and Pkb6 was investigated for the first time. Autophosphorylation was detected only for the Pkb3 catalytic domain.
Subject(s)
Bifidobacterium/enzymology , Protein Serine-Threonine Kinases/isolation & purification , Bifidobacterium/genetics , Catalytic Domain , Culture Techniques , Escherichia coli/cytology , Escherichia coli/genetics , Escherichia coli/growth & development , Inclusion Bodies/genetics , Phosphorylation , Protein Serine-Threonine Kinases/chemistry , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , SolubilityABSTRACT
We report draft genome sequences of two pyrazinamide (PZA)-resistant isolates, Mycobacterium tuberculosis 13-4152 and 13-2459. Isolate 13-4152 is PZA resistant, though it lacks mutations in known genes of PZA resistance. The comparative analysis of these genomes with those stored in GenBank revealed unique mutations, which may elucidate new mechanisms of PZA resistance.
ABSTRACT
The 156 samples of feces separated from healthy residents of the Central region of Russia were used to compose collection of 87 strains of Bifidobacterium out of which 5 strains with wide antagonistic activity related to pathogenic and opportunistic microorganisms were selected. The selected strains are characterized by high probiotic potentials. They have adhesive properties and sensitivity to antibiotics and preparations corresponding to main requirements of common pharmacopoeia articles to strains of microorganisms used in production of probiotics for medicinal application. The given strains of Bifidobacterium can be recommended for development of effective probiotic pharmaceuticals directed to residents of the Central region of Russia.
Subject(s)
Bifidobacterium/isolation & purification , Gastrointestinal Tract/microbiology , Probiotics/metabolism , Adolescent , Adult , Bacillus subtilis/drug effects , Bifidobacterium/classification , Bifidobacterium/metabolism , Candida albicans/drug effects , Feces/microbiology , Female , Humans , Male , Probiotics/pharmacology , Russia , Staphylococcus aureus/drug effects , Young AdultABSTRACT
We report a draft genome sequence of Mycobacterium tuberculosis strain E186hv, belonging to the Beijing B0/W lineage and isolated from a patient from Kurgan, Russia. This clinical isolate showed a reduced virulence phenotype unusual for this lineage and resistance to isoniazid, rifampin, ethambutol, pyrazinamide, and ofloxacin. We analyzed single nucleotide polymorphisms (SNPs) associated with virulence.
ABSTRACT
The patterns of protein phosphorylation in inverted membrane vesicles from the strain Streptomyces fradiae ATCC 19609 were investigated to elucidate the mechanisms of regulation of bacterial membrane bound FoF1-ATP synthase. We found for the first time by two-dimensional gel electrophoresis and mass spectrometry that the ß- and b-subunits of the FoF1-ATP synthase complex undergo phosphorylation; 20 proteins with known functions were identified. All eight subunits of FoF1-ATP synthase, i.e. α, ß, γ, δ, ε, a, b, and c, were cloned into Escherichia coli and expressed as recombinant proteins. Using a crude preparation of serine/threonine protein kinases, we demonstrated the phosphorylation of recombinant γ-, ß-, α- and ε-subunits. The ß-subunit was phosphorylated both as a recombinant protein and in vesicles. Differential phosphorylation of membrane-bound and recombinant proteins can be attributed to different pools of protein kinases in each preparation; in addition, certain steps of FoF1-ATP synthase assembly and function might be accompanied by individual phosphorylation patterns. The structure of the operon containing all subunits and regulatory protein I was identified. The phylogenetic similarity of FoF1-ATP synthase from Streptomyces fradiae ATCC 19609 with the respective proteins in saprophytic and pathogenic (including Mycobacterium tuberculosis) bacteria was investigated. Thus, bacterial serine/threonine protein kinases are important for the regulation of FoF1-ATP synthase. From the practical standpoint, our results provide a basis for designing targeted antibacterial drugs.
Subject(s)
ATP Synthetase Complexes/chemistry , ATP Synthetase Complexes/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Streptomyces/enzymology , ATP Synthetase Complexes/genetics , Bacterial Proteins/genetics , Operon , Phosphorylation , Phylogeny , Protein Subunits/chemistry , Protein Subunits/genetics , Protein Subunits/metabolism , Streptomyces/chemistry , Streptomyces/classification , Streptomyces/geneticsABSTRACT
The problem of Mycobacterium tuberculosis virulence, together with drug resistance, is becoming key for the design of drugs with a new mechanism of action and the production of modern concepts and tuberculosis treatment schemes. The review describes gene complexes and their products, including mycolic acids and global regulatory systems at the level of transcriptional, translational, and post-translational modification, etc. The criteria for selection of virulence/pathogenicity factors that might be used for comparative genomic analysis of strains differing in the degree of virulence were recommended. The experimental approaches and test systems for an adequate estimation of the virulence degree of different strains of M. tuberculosis were analyzed.
Subject(s)
Bacterial Proteins , Gene Expression Regulation, Bacterial/physiology , Mycobacterium tuberculosis , Protein Processing, Post-Translational/physiology , Transcription, Genetic/physiology , Virulence Factors , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , Humans , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/metabolism , Mycobacterium tuberculosis/pathogenicity , Virulence Factors/biosynthesis , Virulence Factors/geneticsABSTRACT
The in silico analysis of 36 sequenced genomes of bacteria of the Bifidobacterium genus determined the presence of 19 genes of toxin-antitoxin (TA) systems that belong to the MazEF and RelBE families, including five mazF and two relE genes that encode toxins and 12 relB genes that encode antitoxins. A high level ofgene (at the level of nucleotide changes) and genomic (presence or absence of genes in distinct genomes) polymorphism in the investigated genes was revealed. The highest level of polymorphism was observed in strains of the Bifidobacterium longum species, primarily in relB1-10 genes. Gene and genomic polymorphism might be used to identify the strain of B. longum species. PCR analysis ofgenomic DNA of 30 bifidobacteria strains belonging to the three species, B. longum, B. adolscentis, and B. bifidum, isolated from the intestinal microbiota of astronauts demonstrated the presence of mazF and relB genes. The observed polymorphism of TA genes indicates the presence of differences in bifidobacteria strains isolated from the intestinal microbiota of astronauts before and after space flight and the control group.
Subject(s)
Antitoxins , Bacterial Toxins , Bifidobacterium/genetics , Antitoxins/genetics , Antitoxins/isolation & purification , Bacterial Toxins/genetics , Bacterial Toxins/isolation & purification , Chromosome Mapping , Humans , Intestines/microbiology , MetagenomeABSTRACT
The toxin-antitoxin gene systems (TASs) are present in the genomes of the overwhelming majority of bacteria and archaea. These systems are involved in various cellular regulatory processes (including stress response), and have not been previously investigated in Lactobacilli. We identified 6 putative TASs with toxins belonging to the MazE and RelE superfamilies (PemK1-Ð1Lrh, PemK2-Ð2Lrh, PemK3-RelB2Lrh, RelE1Lrh, RelB3-RelE3Lrh, and YefM-YoeBLrh) in the genomes of annotated strains of Lactobacillus rhamnosus. PCR analyses revealed that all systems were found in the genomes of 15 strains of L. rhamnosus isolated from humans in central Russia. These strains were highly heterogeneous with respect to the presence of TASs, as well as their nucleotide and amino acid sequences. In three cases, the relE1 genes contained IS3 elements. TAS heterogeneity may be used to reveal inter-genus differences between strains. Cloning of the toxin genes of 3 TASs inhibited Escherichia coli growth, thus confirming their functionality. Cell growth arrest caused by expression of the toxin genes could be reverted by the expression of a cognate antitoxins. Transcription of toxin-antitoxin loci in L. rhamnosus was shown by RT-PCR.
Subject(s)
Antitoxins/chemistry , Antitoxins/genetics , Bacterial Proteins/genetics , Lacticaseibacillus rhamnosus/chemistry , Lacticaseibacillus rhamnosus/genetics , Toxins, Biological/chemistry , Toxins, Biological/genetics , Adult , Amino Acid Sequence , Antitoxins/isolation & purification , Bacterial Proteins/chemistry , Bacterial Proteins/isolation & purification , Base Sequence , Feces/microbiology , Gene Expression Regulation, Bacterial , Humans , Infant , Infant, Newborn , Intestines/microbiology , Polymorphism, Genetic , Russia , Saliva/microbiology , Toxins, Biological/isolation & purificationABSTRACT
This review describes and summarizes the data of ESX secretory system peculiarities characteristic of mainly mycobacteria. This system is involved in the secretion of small proteins of the WXG100 family. Some of these proteins represent virulence factors of Mycobacterium tuberculosis and other pathogenic mycobacteria. The role of these proteins in pathogenesis apparently consists of protecting mycobacteria from lysis in the macrophages that absorb them; the cytolysis of macrophages; and, hence, mycobacterium output into the surrounding tissue. A number of proteins that make up this secretory system are homologs of proteins involved in the conjugation process in saprophytic Mycobacterium smegmatis.