Over the billions of years that bacteria have been around, they have evolved several sophisticated protein secretion nanomachines to deliver toxins, hydrolytic enzymes, and effector proteins into their environments. Of these, the type II secretion system (T2SS) is used by Gram-negative bacteria to export a wide range of folded proteins from the periplasm across the outer membrane. Recent findings have demonstrated that components of the T2SS are localized in mitochondria of some eukaryotic lineages, and their behavior is consistent with the presence of a mitochondrial T2SS-derived system (miT2SS). This review focuses on recent advances in the field and discusses open questions concerning the function and evolution of miT2SSs.

• Klíčová slova
• T2SS, evolution, mitochondria, mitochondrial evolution, protein secretion, protein transport, type II secretion system,
• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• bakteriální sekreční systémy metabolismus   MeSH
• gramnegativní bakterie metabolismus   MeSH
• periplazma metabolismus   MeSH
• sekreční systém typu II * metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• bakteriální proteiny MeSH
• bakteriální sekreční systémy MeSH
• sekreční systém typu II * MeSH

The type 2 secretion system (T2SS) is present in some Gram-negative eubacteria and used to secrete proteins across the outer membrane. Here we report that certain representative heteroloboseans, jakobids, malawimonads and hemimastigotes unexpectedly possess homologues of core T2SS components. We show that at least some of them are present in mitochondria, and their behaviour in biochemical assays is consistent with the presence of a mitochondrial T2SS-derived system (miT2SS). We additionally identified 23 protein families co-occurring with miT2SS in eukaryotes. Seven of these proteins could be directly linked to the core miT2SS by functional data and/or sequence features, whereas others may represent different parts of a broader functional pathway, possibly also involving the peroxisome. Its distribution in eukaryotes and phylogenetic evidence together indicate that the miT2SS-centred pathway is an ancestral eukaryotic trait. Our findings thus have direct implications for the functional properties of the early mitochondrion.

• MeSH
• biologické modely MeSH
• Eukaryota klasifikace   genetika   metabolismus   MeSH
• fylogeneze MeSH
• gramnegativní bakterie klasifikace   genetika   metabolismus   MeSH
• konzervovaná sekvence MeSH
• mitochondriální proteiny klasifikace   genetika   metabolismus   MeSH
• mitochondrie genetika   metabolismus   MeSH
• molekulární evoluce * MeSH
• molekulární modely MeSH
• Naegleria klasifikace   genetika   metabolismus   MeSH
• peroxizomy metabolismus   MeSH
• protozoální proteiny klasifikace   genetika   metabolismus   MeSH
• sekreční systém typu II klasifikace   genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• mitochondriální proteiny MeSH
• protozoální proteiny MeSH
• sekreční systém typu II MeSH

Repeats-in-Toxin (RTX) proteins of Gram-negative bacteria are excreted through the type I secretion system (T1SS) that recognizes non-cleavable C-terminal secretion signals. These are preceded by arrays of glycine and aspartate-rich nonapeptide repeats grouped by four to eight β strands into blocks that fold into calcium-binding parallel β-roll structures. The β-rolls are interspersed by linkers of variable length and sequence and the organization of multiple RTX repeat blocks within large RTX domains remains unknown. Here we examined the structure and function of the RTX domain of Bordetella pertussis adenylate cyclase toxin (CyaA) that is composed of five β-roll RTX blocks. We show that the non-folded RTX repeats maintain the stability of the CyaA polypeptide in the Ca2+-depleted bacterial cytosol and thereby enable its efficient translocation through the T1SS apparatus. The efficacy of secretion of truncated CyaA constructs was dictated by the number of retained RTX repeat blocks and depended on the presence of extracellular Ca2+ ions. We further describe the crystal structure of the RTX blocks IV-V of CyaA (CyaA1372-1681) that consists of a contiguous assembly of two β-rolls that differs substantially from the arrangement of the RTX blocks observed in RTX lipases or other RTX proteins. These results provide a novel structural insight into the architecture of the RTX domains of large RTX proteins and support the "push-ratchet" mechanism of the T1SS-mediated secretion of very large RTX proteins.

• Klíčová slova
• Bordetella pertussis, RTX proteins, adenylate cyclase toxins, type I secretion system,
• MeSH
• adenylátcyklasový toxin chemie   genetika   metabolismus   MeSH
• bakteriální proteiny chemie   metabolismus   MeSH
• bakteriální toxiny chemie   genetika   metabolismus   MeSH
• Bordetella pertussis metabolismus   MeSH
• cytosol metabolismus   MeSH
• gramnegativní bakterie metabolismus   MeSH
• konformace proteinů MeSH
• sbalování proteinů MeSH
• sekreční systém typu I MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenylátcyklasový toxin MeSH
• bakteriální proteiny MeSH
• bakteriální toxiny MeSH
• sekreční systém typu I MeSH

Calcium-binding RTX proteins are equipped with C-terminal secretion signals and translocate from the Ca(2+)-depleted cytosol of Gram-negative bacteria directly into the Ca(2+)-rich external milieu, passing through the "channel-tunnel" ducts of type I secretion systems (T1SSs). Using Bordetella pertussis adenylate cyclase toxin, we solved the structure of an essential C-terminal assembly that caps the RTX domains of RTX family leukotoxins. This is shown to scaffold directional Ca(2+)-dependent folding of the carboxy-proximal RTX repeat blocks into β-rolls. The resulting intramolecular Brownian ratchets then prevent backsliding of translocating RTX proteins in the T1SS conduits and thereby accelerate excretion of very large RTX leukotoxins from bacterial cells by a vectorial "push-ratchet" mechanism. Successive Ca(2+)-dependent and cosecretional acquisition of a functional RTX toxin structure in the course of T1SS-mediated translocation, through RTX domain folding from the C-terminal cap toward the N terminus, sets a paradigm that opens for design of virulence inhibitors of major pathogens.

• MeSH
• adenylátcyklasový toxin chemie   metabolismus   MeSH
• bakteriální toxiny chemie   metabolismus   MeSH
• Bordetella pertussis chemie   enzymologie   MeSH
• buněčné linie MeSH
• gramnegativní bakterie chemie   metabolismus   MeSH
• molekulární modely MeSH
• myši MeSH
• sbalování proteinů MeSH
• sekreční systém typu I metabolismus   MeSH
• sekundární struktura proteinů MeSH
• transport proteinů MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenylátcyklasový toxin MeSH
• bakteriální toxiny MeSH
• sekreční systém typu I MeSH
• vápník MeSH

Outer membrane vesicles secreted by gram-negative bacteria play an important role in bacterial physiology as well as in virulence and host-pathogen interaction. Isolated vesicles of some bacteria have also been studied for their immunomodulatory potential in the vaccine development. However, the production of vesicles in sufficient amount, purity and reproducibility remains a critical challenge for subsequent analyses in most bacteria. In the present review methods of production, isolation, purification and quantification of outer membrane vesicles are summarized and discussed.

• Klíčová slova
• Bacterial secretion, Gram-negative bacteria, Host–pathogen interaction, Outer membrane vesicles, Vaccine development,
• MeSH
• buněčná stěna * imunologie   metabolismus   MeSH
• frakcionace buněk metody   MeSH
• fyziologický stres MeSH
• genetické inženýrství MeSH
• gramnegativní bakterie * genetika   imunologie   metabolismus   MeSH
• sekreční vezikuly * imunologie   metabolismus   MeSH
• subcelulární frakce MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

In this study, 99 Gram-negative rod bacteria were isolated from cooling tower water, and biofilm samples were examined for cell-to-cell signaling systems, N-acyl homoserine lactone (AHL) signal molecule types, and biofilm formation capacity. Four of 39 (10 %) strains isolated from water samples and 14 of 60 (23 %) strains isolated from biofilm samples were found to be producing a variety of AHL signal molecules. It was determined that the AHL signal molecule production ability and the biofilm formation capacity of sessile bacteria is higher than planktonic bacteria, and there was a statistically significant difference between the AHL signal molecule production of these two groups (p < 0.05). In addition, it was found that bacteria belonging to the same species isolated from cooling tower water and biofilm samples produced different types of AHL signal molecules and that there were different types of AHL signal molecules in an AHL extract of bacteria. In the present study, it was observed that different isolates of the same strains did not produce the same AHLs or did not produce AHL molecules, and bacteria known as AHL producers did not produce AHL. These findings suggest that detection of signal molecules in bacteria isolated from cooling towers may contribute to prevention of biofilm formation, elimination of communication among bacteria in water systems, and blockage of quorum-sensing controlled virulence of these bacteria.

• MeSH
• acylbutyrolaktony chemie   izolace a purifikace   metabolismus   MeSH
• biofilmy růst a vývoj   MeSH
• fyziologie bakterií * MeSH
• gramnegativní bakterie izolace a purifikace   metabolismus   MeSH
• mikrobiologie vody * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acylbutyrolaktony MeSH

Formation of biogenic amines may occur in food due to metabolic activities of contaminating Gram-negative bacteria. Putrescine is assumed to be the major biogenic amine associated with microbial food spoilage. Gram-negative bacteria can form putrescine by three metabolic pathways that can include eight different enzymes. The objective of this study was to design new sets of primers able to detect all important enzymes involved in the production of putrescine by Gram-negative bacteria. Seven new sets of consensual primers based on gene sequences of different bacteria were designed and used for detection of the speA, adiA, adi, speB, aguA, speC, and speF genes. A newly developed touchdown polymerase chain reaction (PCR) method using these primers was successfully applied on several putrescine-producers. Selected PCR products were sequenced and high similarity of their sequences (99-91%) with known sequences of the corresponding genes confirmed high specificity of the developed sets of primers. Furthermore, all the investigated bacteria produced both putrescine and agmatine, an intermediate of putrescine production, which was confirmed by chemical analysis. The developed new touchdown PCR method could easily be used to detect potential foodborne Gram-negative producers of putrescine. The newly developed sets of primers could also be useful in further research on putrescine metabolism in contaminating microbiota.

• MeSH
• bakteriální proteiny genetika   MeSH
• DNA primery genetika   MeSH
• gramnegativní bakterie genetika   izolace a purifikace   metabolismus   MeSH
• kontaminace potravin analýza   MeSH
• polymerázová řetězová reakce metody   MeSH
• potravinářská mikrobiologie MeSH
• putrescin biosyntéza   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální proteiny MeSH
• DNA primery MeSH
• putrescin MeSH

Two thermophilic strains belonging to Geobacillus stearothermophilus and Meiothermus ruber, which naturally do not synthesize ω-alicyclic fatty acids (ω-FAs) were cultivated with cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl carboxylic acids. Gas chromatography-mass spectrometry analysis of fatty acid methyl and picolinyl esters showed that both strains are able to synthesize ω-FAs when cultivated with the appropriate precursor. The incorporation of cyclic acids influenced the whole FA composition as well as membrane fluidity. Membrane fluidity of intact cells was studied by measuring the fluorescence polarisation of the probe l,6-diphenyl-1,3,5-hexatriene incorporated into membrane lipid bilayers. Cytoplasmic membrane became more fluid with increasing content of ω-FAs. This is caused by considerable changes in lipid packing within the membrane induced by the presence of ω-FAs not found in the natural environment of Geobacillus and Meiothermus strains.

• MeSH
• buněčná membrána metabolismus   MeSH
• difenylhexatrien metabolismus   MeSH
• fluidita membrány * MeSH
• fluorescenční barviva metabolismus   MeSH
• fluorescenční polarizace MeSH
• Geobacillus stearothermophilus metabolismus   MeSH
• gramnegativní bakterie klasifikace   metabolismus   MeSH
• kyseliny karboxylové metabolismus   MeSH
• nenasycené mastné kyseliny biosyntéza   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• difenylhexatrien MeSH
• fluorescenční barviva MeSH
• kyseliny karboxylové MeSH
• nenasycené mastné kyseliny MeSH

Repeats-in-toxin (RTX) exoproteins of Gram-negative bacteria form a steadily growing family of proteins with diverse biological functions. Their common feature is the unique mode of export across the bacterial envelope via the type I secretion system and the characteristic, typically nonapeptide, glycine- and aspartate-rich repeats binding Ca(2+) ions. In this review, we summarize the current state of knowledge on the organization of rtx loci and on the biological and biochemical activities of therein encoded proteins. Applying several types of bioinformatic screens on the steadily growing set of sequenced bacterial genomes, over 1000 RTX family members were detected, with the biological functions of most of them remaining to be characterized. Activities of the so far characterized RTX family members are then discussed and classified according to functional categories, ranging from the historically first characterized pore-forming RTX leukotoxins, through the large multifunctional enzymatic toxins, bacteriocins, nodulation proteins, surface layer proteins, up to secreted hydrolytic enzymes exhibiting metalloprotease or lipase activities of industrial interest.

• MeSH
• aminokyselinové motivy MeSH
• bakteriální proteiny chemie   genetika   metabolismus   MeSH
• bakteriální toxiny chemie   genetika   metabolismus   MeSH
• gramnegativní bakterie chemie   genetika   metabolismus   MeSH
• multigenová rodina * MeSH
• regulace genové exprese u bakterií MeSH
• transport proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• bakteriální proteiny MeSH
• bakteriální toxiny MeSH

Beta-lactamases are the most common cause of beta-lactam resistance in Gram-negative bacteria. With third-generation and fourth-generation cephalosporins being introduced into practice, new beta-lactamases have evolved, able to hydrolyze these antibiotics. AmpC-type beta lactamases (cephalosporinases) are serine enzymes with the ability to hydrolyze penicillins, monobactams and cephalosporins of all generations, including cephamycins. Over the last two decades, transferable plasmid-mediated class C beta-lactamases have been reported with increasing frequency. The genes for resistance to other groups of antibiotics are usually carried on the same mobile element as the AmpC genes. A reliable method for AmpC detection in routine diagnosis has not been available yet. The issue of AmpC-type beta lactamases is summarized, including their identification, interpretation of susceptibility test results and recommended treatment of infection caused by AmpC producers.

• MeSH
• bakteriální proteiny biosyntéza   genetika   izolace a purifikace   MeSH
• beta-laktamasy biosyntéza   genetika   izolace a purifikace   MeSH
• beta-laktamová rezistence * MeSH
• cefalosporinasa biosyntéza   MeSH
• enzymová indukce MeSH
• gramnegativní bakterie genetika   metabolismus   MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• AmpC beta-lactamases MeSH Prohlížeč
• bakteriální proteiny MeSH
• beta-laktamasy MeSH
• cefalosporinasa MeSH