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J Barbé

Publications and source records attributed to J Barbé.

85 records · Page 5Linked to original sources
PubMed1985

Expression of the SOS genes of Escherichia coli in Salmonella typhimurium.

To lysogenize Salmonella typhimurium by Lambda phage, a region of 10.2 kb of Escherichia coli DNA carrying the nusA gene was cloned in a S. typhimurium strain containing a F'112 plasmid which codifies for the lamB region of E. coli. The strain of S. typhimurium obtained in this way, was lysogenized by lambda c IndO- bacteriophage harboring either a fusion between recA1 or sfiA genes of E. coli with lacZ gene. Likewise, pSE143 plasmid with a umu C::lacZ fusion was introduced in S. typhimurium. Afterwards, induction of these SOS genes was studied. Results obtained show that the basal transcription of both recA and sfiA genes of E. coli was higher in S. typhimurium than in E. coli. Nevertheless, induction of recA and sfiA genes by UV-irradiation and mitomycin C was higher in E. coli than in S. typhimurium. On the other hand, umuC gene of E. coli presents the same basal level of transcription in both E. coli and S. typhimurium species, although induction of this gene by UV-irradiation and mitomycin C was higher in S. typhimurium than in E. coli. Therefore, the plasmid pUA25 constructed in this work may be used to introduce, using the Lambda phage as a vector, the SOS genes of E. coli in other bacterial species which may be useful to study the relationship between their respective SOS systems.

Bacterial Proteins↗
PubMed1984

Effect of L-ethionine on the expression of the SOS system in Escherichia coli.

The effect of L-ethionine, the ethyl analog of the essential amino acid methionine, on the SOS system of Escherichia coli was studied. This compound does not induce either inhibition of cell division nor cessation of cell respiration in a RecA+ Met+ RelA+ strain, nor in RecA+ Met- RelA+ or RecA+ Met- RelA- mutants. Nevertheless, L-ethionine blocks the expression of both cited SOS functions in a recA441 mutant when it is growing at the restrictive temperature of 42 degrees C. Furthermore, the inhibitory effect of the L-ethionine on the induction of the SOS system in this mutant is increased when the cells are preincubated for several hours in the presence of the analog, before the temperature shift. Moreover, cultures of the recA441 mutant incubated at 42 degrees C in the presence of both L-ethionine and L-methionine present the same behaviour as the cultures of this mutant growing at the same temperature but without either amino acid. On the other hand, L-ethionine does not have any effect on the expression of the two mentioned SOS functions when these are induced by UV-irradiation in a RecA+ strain even if this compound is added to the cells several hours before irradiation.

DNA Repair↗
PubMed1984

Changes in ATP concentration in Escherichia coli during induction of the SOS system by mitomycin C and bleomycin.

Treatment of Escherichia coli with bleomycin induced a dramatic increase in ATP concentration in the first 30 min. Afterwards, in RecA+ strains, ATP dropped quickly to values similar to those of untreated cells. Mutants of E. coli defective in either RecA protein or RecA protease activity did not show this decrease, indicating that it was due to the action of RecA protease. The increase in ATP in the first 30 min was dependent on RecBC exonuclease activity and must have been due to substrate level phosphorylation, since an uncoupler such as dinitrophenol did not affect it. Nevertheless, mitomycin C did not induce any change in ATP pools of RecA+ strains, at least during 120 min following treatment. The implications of these findings are discussed in relation to the possible pathways of activation of RecA protease.

Adenosine Triphosphate↗
PubMed1983

Evolution of cellular ATP concentration after UV-mediated induction of SOS system in Escherichia coli.

UV-irradiation of E. coli induces a two fold increase in ATP pool in the first 20 min. Afterwards, in RecA+ strains ATP level drops quickly below values of non irradiated cells. Mutants of E. coli defective in RecA protein or with either RecA protease activity deficient or protease resistant LexA repressor do not present this decrease, showing that it is due to cleavage of LexA repressor by RecA protease. The ATP increase produced in the first 20 min is dependent on RecBC exonuclease activity and it must be due to substrate level phosphorylation since an uncoupler such as dinitrophenol does not affect it.

Adenosine Triphosphate↗
PubMed1983

Indirect induction of SOS functions in Salmonella typhimurium.

Infection of non-UV-irradiated cells of Salmonella typhimurium with UV-damaged P22 or KB1 phage induces recA-dependent inhibition of cell division, cell mutagenesis and prophage induction but not inhibition of respiration. On the contrary, respiration and ATP concentration are increased after treatment with UV-damaged phage in both RecA+ and RecA- strains, showing that this increase is not recA-dependent. Furthermore, infection with UV-damaged phage prevents both inhibition of respiration and decrease in ATP level in the UV-irradiated RecA+ strain. This indirect induction of SOS functions is related to degradation of phage DNA as well as to the multiplicity of infection used, suggesting that DNA degradation may play an important role in the mechanism of expression of the SOS system. Our results give also support to the hypothesis that there exists a differentiation in the expression of the various SOS functions.

Salmonella Phages↗
PubMed1982

Expression of recA-gene dependent SOS functions in Salmonella typhimurium.

Thymine starvation of RecA+ Thy- strains of Salmonella typhimurium does not induce the inhibition of cellular respiration, one of the recA-gene dependent SOS functions. Nevertheless, thymine deprivation is able to produce a normal induction of prophage and thymineless death in these same strains However, when these mutants are treated, in the presence of thymine, with UV-irradiation or bleomycin, they show a normal inhibition of cellular respiration and other SOS functions. Thus, one injurious treatment (thymine deprivation) may trigger prophage induction but not cessation of respiration, whereas another agent (UV-irradiation) may induce both. Together, these results suggest a possible discrimination in the pathways and conditions of expression of various SOS functions.

Bleomycin↗
PubMed1982

A multifunctional gene (tetR) controls Tn10-encoded tetracycline resistance.

The tetracycline resistance regulatory gene (tetR) of transposon Tn10 was analyzed by a combination of methods involving gene fusion and cloning. This gene is located on a 695-base pair HincII DNA fragment near the center of Tn10. The direction of transcription is opposite to that of neighboring gene tetA, which encodes the TetA protein. The gene product of the tetR gene (the TetR protein) has a molecular weight of 23,000. tet-R-lacZ gene fusions encode fusion beta-galactosidases that are membrane bound, indicating that the TetR protein itself is membrane associated. Mutants defective in tetR result in constitutive tetracycline resistance, but the level of resistance is reduced. Expression of the tetR gene is induced by tetracycline; in the absence of tetracycline, the TetR protein turns off its own synthesis.

Bacterial Proteins↗
PubMedDate not supplied

Expression of the SOS response following simultaneous treatment with methyl-nitrosoguanidine and mitomycin C in Escherichia coli.

Simultaneous treatment of Escherichia coli cultures with methyl-nitrosoguanidine and mitomycin C induces recA-dependent inhibition of respiration but not inhibition of cell division. This pattern of SOS functions expression is the same as that is found following treatment with methyl-nitrosoguanidine alone and contrary to the pattern induced after mitomycin C addition. The same result is obtained when a culture of E. coli RecA441 (formerly tif) is shifted to 42 degrees C and treated simultaneously with methyl-nitrosoguanidine. The suppressor effect of this compound over the pattern of SOS functions expression induced by mitomycin C or high temperature in recA441 mutants is directly related to the increase in its dose. Moreover, the division temperature-sensitive mutant ftsA treated with methyl-nitrosoguanidine and high temperature does not show any decrease in its normal filamentous growth when cultured at 42 degrees C. This indicates that the effect of methyl-nitrosoguanidine on the recA-independent inhibition of cell division is not due to any indiscriminate effect of this compound over the division process. These results suggest that the specific kind of lesion caused in DNA is very important in determining which SOS function is induced.

Antibiotics, Antineoplastic↗
PubMedDate not supplied

Molecular fingerprinting of Salmonella typhimurium by IS200-typing as a tool for epidemiological and evolutionary studies.

The aim of this work was to develop and evaluate a molecular typing strategy for Salmonella based on hybridization of chromosomal DNA with two different probes derived from insertion sequence IS200. Probe IS200-TT was specifically constructed for this study as a trimer of a 112 pb TaqI-TaqI fragment of IS200. Among several restriction enzymes evaluated, two were selected: EcoRI, which cuts the insertion sequence in two pieces, each one complementary to one of the probes used, and PstI, a restriction enzyme with no recognition site into IS200. With several combinations of these restrictions enzymes and probes, 43 Salmonella typhimurium strains were analyzed for copy number and location of IS200, as well as reproducibility and stability of the patterns. IS200 types have been shown to be stable, both in strains isolated from different patients implicated in the same salmonellosis outbreak and in strains isolated from the same patient at different times or from different specimens. The discriminatory power of the method has been 0.91 to 0.94. As a comparison, S. typhimurium strains were also ribotyped. Discriminatory power of the ribotypes oscillated between 0.44 and 0.55, depending on the enzyme used, and achieved a 0.78 value when the information obtained with two restriction enzymes was combined. Moreover, IS200 typing was able to differentiate among a group of S. typhimurium strains which were identical by ribotype and enzymatic electrophoretic mobility. These results enable us to conclude that, for the stability, reproducibility and discriminatory power of the patterns generated, IS200 probes can be a very useful tool in the molecular typing of S. typhimurium.

Bacterial Typing Techniques↗