共查询到20条相似文献,搜索用时 15 毫秒
1.
María V. Niklison-Chirou Fernando Dupuy Liliana B. Pena Susana M. Gallego Maria Laura Barreiro-Arcos Cesar Avila Clarisa Torres-Bugeau Beatriz E. Arcuri Augusto Bellomio Carlos Minahk Roberto D. Morero 《The international journal of biochemistry & cell biology》2010,42(2):273-281
We previously showed that the antimicrobial peptide microcin J25 induced the over-production of reactive oxygen species with the concomitant release of cytochrome c from rat heart mitochondria via the opening of the mitochondrial permeability transition pore. Here, we were able to demonstrate that indeed, as a consequence of the oxidative burst, MccJ25 induces carbonylation of mitochondrial proteins, which may explain the irreversible inhibition of complex III and the partial inhibition of superoxide dismutase and catalase. Moreover, the peptide raised the levels of oxidized membrane lipids, which triggers the release of cytochrome c. From in silico analysis, we hypothesize that microcin would elicit these effects through interaction with heme c1 at mitochondrial complex III. On the other hand, under an excess of l-arginine, MccJ25 caused nitric oxide overproduction with no oxidative damage and a marked inhibition in oxygen consumption. Therefore, a beneficial anti-oxidative activity could be favored by the addition of l-arginine. Conversely, MccJ25 pro-oxidative–apoptotic effect can be unleashed in either an arginine-free medium or by suppressing the nitric oxide synthase activity. 相似文献
2.
Sophie Sable Anne-Marie Pons Sandrine Gendron-Gaillard Gilles Cottenceau 《Applied microbiology》2000,66(10):4595-4597
The inhibitory activities of known microcins were evaluated against some diarrheagenic Escherichia coli strains. Some antibacterial properties of microcin J25, the most active one, were studied. A rapid two-step purification was performed. The MIC and the minimum bactericidal concentration of J25 against E. coli O157:H7 were 1 and 100 μg ml−1, respectively. A 104-CFU ml−1 contamination by this strain was destroyed in milk and meat extract by 6.25 μg of J25 ml−1 and in half-diluted egg yolk by 50 μg of J25 ml−1. 相似文献
3.
Niklison Chirou M Bellomio A Dupuy F Arcuri B Minahk C Morero R 《The FEBS journal》2008,275(16):4088-4096
Microcin J25, an antimicrobial lasso-structure peptide, induces the opening of mitochondrial permeability transition pores and the subsequent loss of cytochrome c. The microcin J25 effect is mediated by the stimulation of superoxide anion overproduction. An increased uptake of calcium is also involved in this process. Additional studies with superoxide dismutase, ascorbic acid and different specific inhibitors, such as ruthenium red, cyclosporin A and Mn(2+), allowed us to establish a time sequence of events starting with the binding of microcin J25, followed by superoxide anion overproduction, opening of mitochondrial permeability transition pores, mitochondrial swelling and the concomitant leakage of cytochrome c. 相似文献
4.
Identification and Properties of the Genes Encoding Microcin E492 and Its Immunity Protein 总被引:2,自引:0,他引:2
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
The gene coding for the immunity protein (mceB) and the structural gene of microcin E492 (mceA), a low-molecular-weight channel-forming bacteriocin produced by a strain of Klebsiella pneumoniae, have been characterized. The microcin gene codes for a precursor protein of either 99 or 103 amino acids. Protein sequencing of the N-terminal region of microcin E492 unequivocally identified this gene as the microcin structural gene and indicated that this microcin is synthesized as a precursor protein that is cleaved at either amino acid 15 or 19, at a site resembling the double-glycine motif. The gene encoding the 95-amino-acid immunity protein (mceB) was identified by cloning the DNA segment that encodes only this polypeptide into an expression vector and demonstrating the acquisition of immunity to microcin E492. As expected, the immunity protein was found to be associated with the inner membrane. Analysis of the DNA sequence indicates that these genes belong to the same family as microcin 24, and they do not share structural motifs with any other known channel-forming bacteriocin. The organization of the microcin- and immunity protein-encoding genes suggests that they are coordinately expressed. 相似文献
5.
6.
Andrés Marcoleta Macarena Marín Gabriela Mercado José María Valpuesta Octavio Monasterio Rosalba Lagos 《Journal of bacteriology》2013,195(17):3995-4004
Microcin E492, a channel-forming bacteriocin with the ability to form amyloid fibers, is exported as a mixture of two forms: unmodified (inactive) and posttranslationally modified at the C terminus with a salmochelin-like molecule, which is an essential modification for conferring antibacterial activity. During the stationary phase, the unmodified form accumulates because expression of the maturation genes mceIJ is turned off, and microcin E492 is rapidly inactivated. The aim of this work was to demonstrate that the increase in the proportion of unmodified microcin E492 augments the ability of this bacteriocin to form amyloid fibers, which in turn decreases antibacterial activity. To this end, strains with altered proportions of the two forms were constructed. The increase in the expression of the maturation genes augmented the antibacterial activity during all growth phases and delayed the loss of activity in the stationary phase, while the ability to form amyloid fibers was markedly reduced. Conversely, a higher expression of microcin E492 protein produced concomitant decreases in the levels of the modified form and in antibacterial activity and a substantial increase in the ability to form amyloid fibers. The same morphology for these fibers, including those formed by only the unmodified version, was observed. Moreover, seeds formed using exclusively the nonmodified form were remarkably more efficient in amyloid formation with a shorter lag phase, indicating that the nucleation process is probably improved. Unmodified microcin E492 incorporation into amyloid fibers was kinetically more efficient than the modified form, probably due to the existence of a conformation that favors this process. 相似文献
7.
As Escherichia coli strains enter the stationary phase of growth they become more resistant to the peptide antibiotic microcin J25. It is known that starvation for nutrients such as amino acids or glucose leads to increases in guanosine 3',5'-bispyrophosphate (ppGpp) levels and that the intracellular concentration of this nucleotide increases as cells enter the stationary phase of growth. Therefore, we examined the effects of artificially manipulating the ppGpp levels on sensitivity to microcin J25. A direct correlation was found between ppGpp accumulation and microcin resistance. Our results indicate that the nucleotide is required to induce production of YojI, a chromosomally encoded efflux pump which, in turn, expels microcin from cells. This would maintain the intracellular level of the antibiotic below a toxic level. 相似文献
8.
9.
N E Kurepina I A Khmel' V A Lipasova D A Za?tsev 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1990,(5):12-17
A new microcin produced by an Citrobacter R51 strain has been detected. This antibiotic has been shown to inhibit the growth of a number of Gram-negative as well as Gram-positive strains of bacteria on the minimal medium plates. The properties of partially purified microcin were characterized. Constitutive synthesis of microcin is determined by a conjugative plasmid. The genes of microcin synthesis and immunity were cloned on a plasmid and plasmid vehicles. A physical map of the 12 kb fragment coding for the production of microcin R51 and immunity to this antibiotic is presented. 相似文献
10.
A.E. Galván M.C. Chalón L.A. Schurig-Briccio R.A. Salomón C.J. Minahk R.B. Gennis A. Bellomio 《BBA》2018,1859(2):110-118
Microcin J25 has two targets in sensitive bacteria, the RNA polymerase, and the respiratory chain through inhibition of cellular respiration. In this work, the effect of microcin J25 in E. coli mutants that lack the terminal oxidases cytochrome bd-I and cytochrome bo3 was analyzed. The mutant strains lacking cytochrome bo3 or cytochrome bd-I were less sensitive to the peptide. In membranes obtained from the strain that only expresses cytochrome bd-I a great ROS overproduction was observed in the presence of microcin J25. Nevertheless, the oxygen consumption was less inhibited in this strain, probably because the oxygen is partially reduced to superoxide. There was no overproduction of ROS in membranes isolated from the mutant strain that only express cytochrome bo3 and the inhibition of the cellular respiration was similar to the wild type. It is concluded that both cytochromes bd-I and bo3 are affected by the peptide. The results establish for the first time a relationship between the terminal oxygen reductases and the mechanism of action of microcin J25. 相似文献
11.
María Fernanda Pomares Raúl A. Salomón Olga Pavlova Konstantin Severinov Ricardo Farías Paula A. Vincent 《Applied and environmental microbiology》2009,75(17):5734-5738
Microcin J25 (MccJ25) is a 21-residue ribosomally synthesized lariat peptide antibiotic. MccJ25 is active against such food-borne disease-causing pathogens as Salmonella spp., Shigella spp., and Escherichia coli, including E. coli O157:H7 and non-O157 strains. MccJ25 is highly resistant to digestion by proteolytic enzymes present in the stomach and intestinal contents. MccJ25 would therefore remain active in the gastrointestinal tract, affecting normal intestinal microbiota, and this limits the potential use of MccJ25 as a food preservative. In the present paper, we describe a chymotrypsin-susceptible MccJ25 derivative with a mutation of Gly12 to Tyr that retained almost full antibiotic activity and efficiently inhibited the growth of pathogenic Salmonella enterica serovar Newport and Escherichia coli O157:H7 in skim milk and egg yolk. However, unlike the wild-type MccJ25, the MccJ25(G12Y) variant was inactivated by digestive enzymes both in vitro and in vivo. To our knowledge, our results represent the first example of a rational modification of a microcin aimed at increasing its potential use in food preservation.Escherichia coli microcin J25 (MccJ25) is a plasmid-encoded antibiotic peptide consisting of 21 amino acid residues (G1-G-A-G-H5-V-P-E-Y-F10-V-G-I-G-T15-P-I-S-F-Y20-G) (4, 12). Four genes (mcjA, mcjB, mcjC, and mcjD) are required for MccJ25 synthesis, export, and immunity (14, 15). The mcjA gene encodes a 58-amino-acid MccJ25 precursor, which is processed by the products of mcjC and mcjB (7). Once synthesized, the mature MccJ25 is excreted to the medium by McjD, an ABC-type transporter (6, 14). The tertiary structure of MccJ25 was elucidated as a lariat peptide (1, 10, 17). It contains an eight-residue ring (G1 to E8) and a tail (Y9 to G21) whose C-terminal end is sterically trapped within the ring. MccJ25 amino acids F10 to P16 form a β-hairpin structure, comprising two β-strands (F10-V11 and T15-P16) and a β-turn (V11 to G14).MccJ25 is active on gram-negative bacteria related to the producer strain, and among them are several human pathogens (11, 12, 16). It was previously shown that the E. coli RNA polymerase (5, 18) and the bacterial respiratory chain (2, 9) are the targets for MccJ25 action. MccJ25 is active on pathogenic strains of Salmonella spp., Shigella spp. (12), and E. coli, including O157:H7 (11) as well as non-O157 strains (data not shown), that frequently cause outbreaks of food-borne diseases. In addition, Sable et al. (11) showed that MccJ25 was the most active microcin against 12 out of 15 diarrheagenic E. coli strains tested. These authors also demonstrated that MccJ25 inhibits E. coli O157:H7 in biological products such as milk, egg yolk, and meat extract. These findings suggest that MccJ25 could be an efficient complement to nisin for food preservation. However, the potential usefulness of MccJ25 is compromised by the fact that it is highly resistant to digestion by proteolytic enzymes of the stomach (pepsin) and intestinal (trypsin, chymotrypsin, and carboxypeptidases) contents. Thus, the antibiotic would most likely remain active in the intestine, and this could lead to disturbance of the normal microbiota. Therefore, for potential use of MccJ25 as a food additive, it would be desirable to render MccJ25 susceptible to at least one of these proteases. In the present work, we describe a chymotrypsin-susceptible MccJ25 derivative that remains fully active on S. Newport and E. coli O157:H7 in biological products, namely milk and egg yolk. In addition, we demonstrate that the peptide is inactivated by rat intestinal contents. 相似文献
12.
Laila Ben Said Jean-Guillaume Emond-Rheault Samira Soltani Sofiane Telhig Séverine Zirah Sylvie Rebuffat Moussa Sory Diarra Lawrence Goodridge Roger C. Levesque Ismail Fliss 《Environmental microbiology》2020,22(7):2907-2920
In livestock production, antibiotics are used to promote animal growth, control infections and thereby increase profitability. This practice has led to the emergence of multiresistant bacteria such as Salmonella, of which some serovars are disseminated in the environment. The objective of this study is to evaluate microcin J25 as an inhibitor of Salmonella enterica serovars of various origins including human, livestock and food. Among the 116 isolates tested, 37 (31.8%) were found resistant to at least one antibiotic, and 28 were multiresistant with 19 expressing the penta-resistant phenotype ACSSuT. Microcin J25 inhibited all isolates, with minimal inhibitory concentration values ranging from 0.06 μg/ml (28.4 nM) to 400 μg/ml (189 μM). Interestingly, no cross-resistance was found between microcin J25 and antibiotics. Multiple sequence alignments of genes encoding for the different proteins involved in the recognition and transport of microcin J25 showed that only ferric-hydroxamate uptake is an essential determinant for susceptibility of S. enterica to microcin J25. Examination of Salmonella strains exposed to microcin J25 by transmission electronic microscopy showed for the first-time involvement of a pore formation mechanism. Microcin J25 was a strong inhibitor of several multiresistant isolates of Salmonella and may have a great potential as an alternative to antibiotics. 相似文献
13.
The interaction of the tryptophan-containing variant of microcin J25, MccJ25 I13W, with phosphatidylcholine membranes was studied by fluorescence spectroscopy techniques. The peptide was able to interact with dimiristoylphophatidylcholine and dipalmitoylphosphatidylcholine liposomes only when the membranes were in gel phase, as was demonstrated by the blue shift of the intrinsic fluorescence of MccJ25 I13W. The binding isotherm showed a cooperative partition of the peptide toward the membrane and the binding constant increased as the temperature decreased and the order parameter increased. No interaction with liquid crystalline membranes was observed. Studies of dynamic quenching of the fluorescence indicated that the peptide penetrated the lipid bilayer and was located primarily in the interfacial region. Our results suggest that MccJ25 I13W interacts with gel phase phospholipids and increases both its own affinity for the bilayer and the membrane permeability of small ions. 相似文献
14.
Niklison Chirou MV Minahk CJ Morero RD 《Biochemical and biophysical research communications》2004,317(3):882-886
In this report we studied the effect of the antimicrobial peptide, microcin J25, on the rat heart mitochondria. This peptide induced an important inhibition of the ATP synthesis with the concomitant enhancement of the ATP degradation. These effects were the result of two processes: on one hand, microcin J25 was able to insert into the membrane and hence alter its permeability with the consequent dissipation of the proton motive force. On the other, microcin J25 inhibited the enzymatic activity of the cytochrome c reductase (complex III) of the respiratory chain. The relevance of this study to the potential use of microcin J25 as an anti-tumoral agent is discussed. 相似文献
15.
In the present study, we showed that yojI, an Escherichia coli open reading frame with an unknown function, mediates resistance to the peptide antibiotic microcin J25 when it is expressed from a multicopy vector. Disruption of the single chromosomal copy of yojI increased sensitivity of cells to microcin J25. The YojI protein was previously assumed to be an ATP-binding-cassette-type exporter on the basis of sequence similarities. We demonstrate that YojI is capable of pumping out microcin molecules. Thus, one obvious explanation for the protective effect against microcin J25 is that YojI action keeps the intracellular concentration of the peptide below a toxic level. The outer membrane protein TolC in addition to YojI is required for export of microcin J25 out of the cell. Microcin J25 is thus the first known substrate for YojI. 相似文献
16.
Microcin J25 is a 2,107-Da, plasmid-encoded, cyclopeptide antibiotic produced by Escherichia coli. We have isolated lacZ fusions to mcjA (encoding the 58-amino-acid microcin precursor) and mcjB and mcjC (which are required for microcin maturation), and the regulation of these fusions was used to identify factors that control the expression of these genes. The mcjA gene was found to be dramatically induced as cells entered the stationary phase. Expression of mcjA could be induced by resuspending uninduced exponential-phase cells in spent supernatant obtained from an early-stationary-phase culture. Induction of mcjA expression was not dependent on high cell density, pH changes, anaerobiosis, or the buildup of some inducer. A starvation for carbon and inorganic phosphate induced mcjA expression, while under nitrogen limitation there was no induction at all. These results taken together suggest that stationary-phase induction of mcjA is triggered by nutrient depletion. The mcjB and mcjC genes were also regulated by the growth phase of the culture, but in contrast to mcjA, they showed substantial expression already during exponential growth. Induction of the microcin genes was demonstrated to be independent of RpoS, the cyclic AMP-Crp complex, OmpR, and H-NS. Instead, we found that the growth-phase-dependent expression of mcjA, mcjB, and mcjC may be explained by the concerted action of the positively acting transition state regulators ppGpp, Lrp, and integration host factor. Measurements of microcin J25 production by strains defective in these global regulators showed a good correlation with the reduced expression of the fusions in such mutant backgrounds. 相似文献
17.
Microcin 25, a peptide antibiotic excreted by an Escherichia coli strain isolated from human feces, was purified to homogeneity and characterized. Composition analysis and data from gel filtration indicated that microcin 25 may contain 20 amino acid residues. It has a blocked amino-terminal end. Microcin synthesis and immunity are plasmid determined, and the antibiotic was produced in minimal medium when the cultures entered the stationary phase of growth. The peptide appears to interfere with cell division, since susceptible cells filamented when exposed to it. This response does not seem to be mediated by the SOS system. 相似文献
18.
Microcin J25 has dual and independent mechanisms of action in Escherichia coli: RNA polymerase inhibition and increased superoxide production
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Bellomio A Vincent PA de Arcuri BF Farías RN Morero RD 《Journal of bacteriology》2007,189(11):4180-4186
Microcin J25 (MccJ25) uptake by Escherichia coli requires the outer membrane receptor FhuA and the inner membrane proteins TonB, ExbD, ExbB, and SbmA. MccJ25 appears to have two intracellular targets: (i) RNA polymerase (RNAP), which has been described in E. coli and Salmonella enterica serovars, and (ii) the respiratory chain, reported only in S. enterica serovars. In the current study, it is shown that the observed difference between the actions of microcin on the respiratory chain in E. coli and S. enterica is due to the relatively low microcin uptake via the chromosomally encoded FhuA. Higher expression by a plasmid-encoded FhuA allowed greater uptake of MccJ25 by E. coli strains and the consequent inhibition of oxygen consumption. The two mechanisms, inhibition of RNAP and oxygen consumption, are independent of each other. Further analysis revealed for the first time that MccJ25 stimulates the production of reactive oxygen species (O(2)(*-)) in bacterial cells, which could be the main reason for the damage produced on the membrane respiratory chain. 相似文献
19.
Microcin B17 blocks DNA replication and induces the SOS system in Escherichia coli 总被引:13,自引:0,他引:13
Microcin B17 is a novel peptide antibiotic of low Mr (about 4000) produced by Escherichia coli strains carrying plasmid pMccB17. The action of this microcin in sensitive cells is essentially irreversible, follows single-hit kinetics, and leads to an abrupt arrest of DNA replication and, consequently, to the induction of the SOS response. RecA- and RecBC- strains are hypersensitive to microcin B17. Strains producing a non-cleavable SOS repressor (lexAl mutant) are also more sensitive than wild-type, whereas strains carrying a mutation which causes constitutive expression of the SOS response (spr-55) are less sensitive to microcin. Microcin B17 does not induce the SOS response in cells which do not have an active replication fork. The results suggest that the mode of action of this microcin is different from all other well-characterized microcins and colicins, and from other antibiotics which inhibit DNA replication. 相似文献
20.
Microcin J25 uptake: His5 of the MccJ25 lariat ring is involved in interaction with the inner membrane MccJ25 transporter protein SbmA
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
de Cristóbal RE Solbiati JO Zenoff AM Vincent PA Salomón RA Yuzenkova J Severinov K Farías RN 《Journal of bacteriology》2006,188(9):3324-3328
Escherichia coli microcin J25 (MccJ25) is a plasmid-encoded antibiotic peptide consisting of 21 L-amino acid residues (G1-G-A-G-H5-V-P-E-Y-F10-V-G-I-G-T15-P-I-S-F-Y20-G). E. coli RNA polymerase (RNAP) is the intracellular target of MccJ25. MccJ25 enters cells after binding to specific membrane transporters: FhuA in the outer membrane and SbmA in the inner membrane. Here, we studied MccJ25 mutants carrying a substitution of His5 by Lys, Arg, or Ala. The inhibitory effects on cellular growth and in vitro RNAP activity were determined for each mutant microcin. The results show that all mutants inhibited RNAP in vitro. However, the mutants were defective in their ability to inhibit cellular growth. Experiments in which the FhuA protein was bypassed showed that substitutions of MccJ25 His5 affected the SbmA-dependent transport. Our results thus suggest that MccJ25 His5 located in the lariat ring is involved, directly or indirectly, in specific interaction with SbmA and is not required for MccJ25 inhibition of RNAP. 相似文献