Biomedical subjects

J Barbé

Publications and source records attributed to J Barbé.

At least 73 records · Page 4Linked to original sources

PubMed1988

Increase in plasmid transformation efficiency in SOS-induced Escherichia coli cells.

UV irradiation of competent cells of Escherichia coli K12 produced an increase in the efficiency of transformation with plasmid DNA. This phenomenon has been called IPTE (increase in plasmid transformation efficiency) and is dependent on the activated state of the RecA protein. IPTE is independent of the lexA, recB recC, and recF genes. It is not related to the size or the replicon type of the plasmid. Furthermore, it is also induced in cells which have been previously treated with other SOS system-inducing agents such as bleomycin, mitomycin C, or nalidixic acid. IPTE is therefore similar to other repair (SOS) functions inducible by DNA damage since all of them are dependent upon activation of the RecA protein. IPTE differs from other SOS functions in the absence of a direct control by the LexA repressor.

DNA Repair↗

PubMed1988

Evidence for a specific regulation of recA gene transcription in Escherichia coli.

The kinetics of the recA, sfiA and umuDC genes transcription were studied during a double SOS-inducing treatment in Escherichia coli cells using several strains carrying lacZ gene fusions. A transient inhibition in recA, but not in sfiA or umuDC promoted beta-galactosidase synthesis was detected after successive UV-irradiations. Results obtained with a recA--lacZ fusion introduced in several DNA-repair mutants demonstrated that neither a lower LexA inactivation nor a decrease in the production of the inducing signal are the events through which the successive UV-irradiation promoted the arrest of recA transcription. On the contrary, a specific UV-dose-dependent delay appears to be the reason for the inhibition of the recA gene transcription in cells irradiated twice.

Adenosine Triphosphate↗

PubMed1988

Adaptive response to simple alkylating agents in the phototrophic bacteria Rhodobacter capsulatus and R.sphaeroides.

The presence of an adaptive response to low doses of alkylating chemicals in the phototrophic bacteria Rhodobacter sphaeroides and R.capsulatus has been studied. Results obtained show that both strains display this repair response against the challenge doses of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), when they are pretreated with low doses of this compound for 120 min. The adaptive response of both R.sphaeroides and R.capsulatus induced an increase of cell survival and a decrease of mutagenesis in the MNNG-pretreated cells. Furthermore, the MNNG-induced adaptive repair also gives protection to diethylsulphate and ethylmethanesulphonate in both phototrophic bacteria. Finally, the MNNG-promoted adaptive response is sensitive to inhibitors of protein synthesis such as chloramphenicol, indicating that this DNA repair mechanism needs protein synthesis in R.sphaeroides and R.capsulatus, in a way similar to that which occurs in Escherichia coli.

Chloramphenicol↗

PubMed1988

Regulation of ubiG gene expression in Escherichia coli.

To study the regulation of the expression in Escherichia coli of the ubiG gene, which codes for the last enzyme in the pathway of ubiquinone biosynthesis, a fusion between the ubiG and lacZ genes was constructed in vitro. The results showed that (i) the expression of the ubiG gene was higher under aerobic conditions than under anaerobic growth conditions, (ii) the presence of glucose in the culture medium decreased the transcription of the ubiG gene, and (iii) cya and crp mutants exhibited lower levels of ubiG gene expression than the wild-type strain. The addition of cyclic AMP increased the expression of the ubiG gene in both cya and wild-type strains but not in a crp mutant. This fact suggests that the cyclic AMP receptor protein-cyclic AMP complex positively modulates ubiG gene transcription. It was also determined that the transcription of the ubiG gene was in the counterclockwise direction on the E. coli map.

Carbon↗

PubMed1988

Inhibition of the SOS response of Escherichia coli by the Ada protein.

Induction of the adaptive response by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) caused a decrease in the UV-mediated expression of both recA and sfiA genes but not of the umuDC gene. On the other hand, the adaptive response did not affect the temperature-promoted induction of SOS response in a RecA441 mutant. The inhibitory effect on the UV-triggered expression of the recA and sfiA genes was not dependent on either the alkA gene or the basal level of RecA protein, but rather required the ada gene. Furthermore, an increase in the level of the Ada protein, caused by the runaway plasmid pYN3059 in which the ada gene is regulated by the lac promoter, inhibited UV-mediated recA gene expression even in cells to which the MNNG-adaptive treatment had not been applied. This inhibitory effect of the adaptive pretreatment was not observed either in RecBC- strains or in RecBC mutants lacking exonuclease V-related nuclease activity. However, RecF- mutants showed an adaptive response-mediated decrease in UV-promoted induction of the recA gene.

Alkylation↗

PubMed1987

Induction of the SOS response by hydroxyurea in Escherichia coli K12.

Hydroxyurea at concentrations higher than 10(-2) M induced the recA and sfiA genes of E. coli as well as the lambda prophage by a pathway independent of the recBC genes. In addition, the hydroxyurea-mediated induction of the SOS response is accompanied by a recA-dependent decrease on the cellular ATP pool. The presence of the multicopy plasmid pPS2, harboring the nrdAB genes (encoding the ribonucleoside reductase enzyme), abolished the hydroxyurea-induced expression of the recA gene. These data lead us to suggest that induction of the SOS response by hydroxyurea is due to the blocking of DNA replication by the inhibition of the ribonucleoside reductase complex activity.

Bacterial Proteins↗

PubMed1987

Influence of S9 mix in the induction of SOS system by quercetin.

The induction of recA, umuC and sfiA genes by quercetin was studied in the presence and in the absence of S9 mix. The inducing activity of quercetin is higher for sfiA than for recA and umuC genes in the absence of S9 mix. The putative genotoxic metabolites of quercetin produced by S9 mix display different inducing activities of the three SOS genes as compared to quercetin. The induction of sfiA gene is decreased by the presence of S9 mix, whereas an opposite effect was observed concerning umuC and recA. These data suggest that the error-prone repair pathway participates in mutagenesis by quercetin and its metabolites. Moreover, the type of DNA damage exerted by quercetin seems to be determined by its metabolic fate. The importance of testing for the induction of other SOS genes, together with sfiA, in the study of SOS functions as a genotoxic index is emphasized.

Animals↗

PubMed1987

DNA repair systems in the phototrophic bacterium Rhodobacter capsulatus.

UV irradiation and mitomycin C exposure trigger a protease-activity-dependent inhibition of cell division in Rhodobacter capsulatus, which begins about 2 h after the treatment is applied. UV irradiation also induces a dose-dependent mutagenesis with a maximal rate between 5 and 10 J m-2, with increased synthesis of a protein of Mr approximately 30,000 between 2 and 3 h after UV irradiation. In addition, R. capsulatus has an efficient photoreactivation system that reverses the lethal effects of UV irradiation in the presence of intense visible light.

Cell Division↗

PubMed1987

Modulating action of cyclic AMP on the expression of two SOS genes in Escherichia coli K-12.

Ultraviolet irradiation and cyclic AMP treatment produce a synergistic effect on the induction of the cle1 gene (coding for bacteriocin ColE1) in wild-type strains of Escherichia coli. On the other hand, cyclic AMP does not affect the uv-mediated induction of the recA, sfiA, and umuDC genes. Growth in the presence of glucose or glycerol does not affect the factor of amplification of the expression of the cle1 gene in uv-irradiated cells of the wild-type strain. Although, in cultures not treated with uv, the basal level of cle1 induction is about twice as high in cell grown grown with glycerol as in those using glucose as carbon source. In recA mutants neither simultaneous nor separate treatments with either cyclic AMP or uv irradiation induced transcription of the cle1 gene. Moreover, cyclic AMP induced a slight increase in cle1 gene expression in uv-irradiated cya strains, but not in the crp mutants. Nevertheless, the pattern of the uv-mediated induction of other SOS genes, such as umuDC, was the same in the cya and crp mutants, as in their parental wild-type strains. Furthermore, the uv-mediated induction of lambda prophage was decreased after either addition of cyclic AMP or growth in cultural conditions where the level of this nucleotide was low.

Bacteriophage lambda↗

PubMed1986

Induction of SOS genes of Escherichia coli by chromium compounds.

The induction of several SOS genes of Escherichia coli such as recA, umuC, and sfiA by hexavalent (K2Cr2O7, K2CrO4, and CrO3) and trivalent (CrCl3, Cr(NO3)3, and (CH3COO)3Cr) compounds of chromium was studied. Induction was measured as beta-galactosidase activity, using lacZ gene fusions under the control region of different SOS genes. The hexavalent chromium forms induced the genes responsible for massive synthesis of RecA protein, error-prone repair, and inhibition of cell division. On the other hand, the trivalent chromium compounds were unable to induce any of the SOS genes tested. Individual assay of hexavalent chromium compounds showed that K2Cr2O7 was a stronger inducing agent of those three SOS genes tested than K2CrO4, which, in turn, was stronger than CrO3. All this data led to the conclusion that hexavalent chromium compounds, but not trivalent, are proficient agents of induction of the SOS system and can produce indirect mutagenesis in Escherichia coli.

Cell Division↗

PubMed1986

Expression of the SOS system in Escherichia coli growing under nitrate respiration conditions.

Induction of several SOS functions by mitomycin C, bleomycin or thermal treatment of a recA441 mutant growing under nitrate respiration conditions was studied in Escherichia coli. Mitomycin C caused inhibition of cell division, induction of prophages and expression of umuC gene but like in aerobically growing cells, it did not trigger the cessation of cell respiration. On the contrary, both recA+ and recA441 cultures either treated with bleomycin or incubated at 42 degrees C failed to induce any of the different SOS functions cited above. Furthermore, after bleomycin addition or thermal treatment both recA+ and recA441 cultures did not present any variation in the cellular ATP level, contrary to what happens under aerobic growth. The blocking of the expression of some SOS functions under nitrate respiration conditions is not an irreversible process because cells incubated under these anaerobic conditions were able to induce the SOS system when changed to an aerobic medium 30 min after the SOS-inducing treatment had been applied.

Adenosine Triphosphate↗

PubMed1986

5-Azacytidine: survival and induction of the SOS response in Escherichia coli K-12.

Survival and induction of the SOS system by 5-azacytidine, an analog of cytidine, were studied in Escherichia coli K-12. This compound did not produce any effect on the viability of dcm and dam dcm mutants. Furthermore, recA430 and lexA1 strains (both mutations interfere with LexA repressor cleavage but not recombination proficiency) were more resistant than the wild-type strain of E. coli K-12. In contrast, recBC and recA13 mutants were more sensitive to 5-azacytidine than the wild type. Transient exposure of E. coli to 5-azacytidine for 60 min induced both recA-dependent inhibition of cell division and induction of lambda prophage in Dcm+ strains but not in Dcm- mutants. Expression of both functions was dependent on recBC exonuclease. On the other hand, 5-azacytidine was unable to trigger the induction of umuCD and mucB genes and no amplification of RecA protein synthesis in either Dcm+ or Dcm- strains was observed. These last results are in agreement with previously reported data suggesting that there is a discrimination in the expression of the several SOS functions and that some SOS genes may be induced without amplification of RecA protein synthesis.

Adenosine Triphosphate↗

PubMed1986

Effect of P22-mediated receptor release and of phage DNA injection on cell viability of Salmonella typhimurium.

Infection with u.v.-inactivated P22 bacteriophage at multiplicities higher than 30 caused a decrease in Salmonella typhimurium viability without cell lysis. Neither the action of the endoglycosidase of the P22 virion on the lipopolysaccharide of S. typhimurium nor the concomitant release of cell wall components was responsible for m.o.i.-dependent cell death. Using both free P22 tails and u.v.-inactivated P22, we have shown that p9 tail protein activity has no effect either on the integrity of host cells or on cell viability. Our results show that cell death is due to the injection of the u.v.-inactivated P22 DNA.

Adsorption↗

PubMed1986

Effect of alkylating agents on the expression of inducible genes of Escherichia coli.

Increasing doses of alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine, diethyl sulphate and ethylmethane sulphonate cause an inhibition of the expression of the recA and sfiA genes of wild-type Escherichia coli. This behaviour was not observed in a lexA56 mutant which has a defective LexA repressor that is unable to bind to the SOS operator. Furthermore, an ada-1 mutant showed the same behaviour as the wild-type strain indicating that the adaptive proteins are not responsible for the inhibition of recA and sfiA at high doses of alkylating agents. These results suggest that the inhibitory effect of these alkylating agents may be found in the interaction between the LexA repressor and the control regions of sfiA and recA. On the other hand, high doses of either UV light or mitomycin C produced only a slight decrease in the induction of recA and sfiA, whereas bleomycin had no effect. The fact that a repressor structurally related to LexA repressor, such as LacI protein, showed the same behaviour as the LexA repressor when a Lac+ strain was treated with alkylating agents, suggests that these compounds can modify the binding abilities of repressors to DNA, producing a limited or even abolished release of repressors, and so decreasing the expression of inducible genes.

Alkylating Agents↗

PubMed1986

Characterization of SE1, a new general transducing phage of Salmonella typhimurium.

A transducing phage, SE1, which is able to infect Salmonella typhimurium was isolated from a Salmonella enteritidis strain. SE1 is a temperate phage which is heteroimmune with respect to phages P22, L, KB1 and ES18. It is similar in morphology and size to phages P22, L and KB1 and is serologically related to phages P22 and L but not to KB1. Efficiencies of generalized transduction effected by phage SE1 are similar to those for P22HT (int7), a mutant which mediates a high frequency of chromosomal gene transduction. The lengths of chromosomal DNA transduced by SE1 and P22HT (int7) are similar. Furthermore, the SE1 prophage does not exclude the transducing particles from cells it has lysogenized; consequently it is possible to use both SE1 lysogens and non-lysogenic strains as recipients in SE1-mediated transduction experiments, and obtain similar transduction efficiencies. However, the SE1 prophage gives rise to a lysogenic conversion that decreases the rate of adsorption of SE1 and L phages by about 50%, but does not affect adsorption of P22. Altogether these results suggest that phage SE1 may be a useful tool in the genetic manipulation of S. typhimurium.

Antigens, Bacterial↗

PubMed1986

ATP hydrolysis during SOS induction in Escherichia coli.

Changes in cellular ATP concentration during SOS induction in strains of Escherichia coli with different levels of RecA and LexA proteins were studied. UV irradiation of RecA+ strains induced a twofold increase in the ATP concentration around the first 20 min, followed by a decrease to the values of nonirradiated cells. On the other hand, mutants defective in RecA protein or with either deficient RecA protease activity or cleavage-resistant LexA repressor did not show any decrease, suggesting that ATP consumption is related to LexA repressor hydrolysis. Furthermore, strains presenting a constitutive synthesis of RecA protein showed the same changes in ATP concentration as the wild-type strain. Likewise, the presence in a RecA+ strain of a LexA(Def) protein, which is defective in its capacity for binding specifically to SOS operators, did not disturb the changes in ATP when compared with the LexA+ RecA+ strain. Moreover, after UV irradiation, a LexA(Def) RecA- double mutant showed an important increase in ATP concentration, which remained elevated for at least 120 min after UV treatment.

Adenosine Triphosphate↗

PubMed1985

Further characterization of SOS system induction in recBC mutants of Escherichia coli.

Expression of several SOS functions such as induction of lambda prophage, inhibition of cell division and induction of both umuC and recA genes after UV-irradiation, nalidixic acid or mitomycin C addition was studied in an RecBC- mutant. UV-irradiation and mitomycin C induced all SOS functions studied in the RecBC- cells but at a lower level and delayed with respect to the wild-type strain. On the contrary, nalidixic acid was unable to trigger any of these SOS functions. In the RecBC- mutant, adenine only had a stimulating effect on the amplification of RecA protein synthesis following UV-irradiation. Nevertheless, in the wild-type strain the stimulating effect occurred in all SOS functions studied following UV-irradiation as well as in the amplification of RecA protein synthesis by nalidixic acid but not in the other SOS functions triggered by this compound. Furthermore, adenine produced a decrease in the mitomycin C-mediated induction of all SOS functions studied in both RecBC- and wild-type strains.

Adenine↗

PubMed1985

Effect of adenine, cytidine and guanosine on the expression of the SOS system in Escherichia coli.

Addition of cytidine or guanosine to UV-irradiated cells of a RecA+ strain of Escherichia coli did not produce any effect on the induction of two SOS functions: inhibition of cell division and expression of the umuC gene. Under the same conditions adenine gave a slight increase in the induction of these two responses. In a RecA441 mutant growing at 42 degrees C, both cytidine and guanosine inhibited these SOS functions, whereas adenine produced a large increase in their expression. Moreover, the ATP concentration of the RecA441 mutant at 42 degrees C showed a decrease which occurred earlier in the cells growing in the presence of cytidine or guanosine than in the absence of either compound. Adenine induced an increase of about three times the initial ATP concentration of this mutant at 42 degrees C which dropped quickly after 10 min. Neither cytidine nor guanosine increased the evolution of cellular ATP in UV-irradiated cells of the RecA+ strain, whereas adenine had only a slight positive effect. However, in UV-irradiated RecA+ cells with and without adenine, ATP levels dropped quickly to the initial value after 20 min. These data suggest that the influence of adenine, cytidine and guanosine on the expression of the RecA441 phenotype at 42 degrees C may be due to alteration of the cellular ATP concentration of this mutant.

Adenine↗