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M Frenette

Publications and source records attributed to M Frenette.

36 records · Page 2Linked to original sources
PubMed1993

Altered expression of several genes in IIIManL-defective mutants of Streptococcus salivarius demonstrated by two-dimensional gel electrophoresis of cytoplasmic proteins.

Mannose, glucose and fructose are transported in Streptococcus salivarius by a phosphoenolpyruvate:mannose phosphotransferase system (PTS) which consists of a membrane-bound Enzyme II (EII) and two forms of IIIMan having molecular weights of 38,900 (IIIManH) and 35,200 (IIIManL), respectively. We have previously reported the isolation of spontaneous mutants lacking IIIManL and showed that they exhibit higher beta-galactosidase activity than the parental strain after growth on glucose, and that some of them constitutively express a fructose PTS which is induced by fructose in the parental strain. In an attempt to determine whether the expression of other genes is affected by the mutation and what the physiological link is between them, we examined three S. salivarius IIIManL-defective mutants (strains A37, B31 and G29) and the parental strain using two-dimensional gel electrophoresis after growth of the cells on a variety of sugars. After growth on glucose, five new proteins were detected in the cytoplasm of the three mutants. Two of these proteins were induced in the parental strain by galactose or oligosaccharides containing galactose, and one was specifically induced by melibiose. The other two proteins were not detected in the parental strain under any of the growth conditions tested. Two other proteins were only detected in glucose-grown cells of mutant A37, and a protein associated with the metabolism of fructose was constitutively expressed in mutants B31 and G29. Moreover, we have found that under identical growth conditions the amounts of several other proteins which were detected in the parental strain were either increased or decreased in the mutants. Globally, our results have indicated that (1) the expression of several genes was affected in the spontaneous IIIManL-defective mutants; (2) some of the proteins abnormally produced in the mutants were specifically induced in the parental strain by sugars; (3) the phenotypic modifications observed in the mutants were of two types: most were observed solely after growth of the cells on glucose whereas the others were glucose-independent; and (4) the mutants shared common phenotypic traits, but also exhibited idiosyncratic characteristics.

Bacterial Proteins↗
PubMed1993

Alterations in the cellular envelope of spontaneous IIIManL-defective mutants of Streptococcus salivarius.

In Streptococcus salivarius, the phosphoenolpyruvate: mannose phosphotransferase system (PTSMan) transports and concomitantly phosphorylates mannose, glucose, fructose and 2-deoxyglucose. PTSMan consists of a membrane Enzyme II and two forms of Enzyme III (IIIMan) having molecular masses of 38.9 kDa (IIIManH) and 35.2 kDa (IIIManL) respectively. We have previously reported the isolation of spontaneous mutants lacking IIIManL, and showed that they exhibited abnormal growth when cultured in mixtures of sugars containing glucose. The mutants also synthesize several cytoplasmic glucose-repressible proteins during growth on glucose and some of them constitutively express a fructose PTS which is induced by fructose in the parental strain. We have now investigated the properties and composition of the cellular envelope of three S. salivarius IIIManL-defective mutants (strains A37, B31 and G29) after growth on glucose. The mutants have altered sensitivity to various toxic compounds that interfere with cell-envelope functions. The mutants also exhibited altered membrane-protein profiles when analysed by two-dimensional PAGE and modified total lipid and phosphorus contents and lipid/protein ratio. In one mutant (strain G29), the proportion of the phospholipids separated by TLC was different from the parental strain. Electron microscopy indicated that one mutant (strain A37) possessed more fimbriae than the parental strain. The results suggested that these IIIManL-defective mutants were affected in a global regulatory gene controlling several cellular or physiological functions, many of these being related to the cellular envelope.

Bacterial Proteins↗
PubMed1992

Cloning, sequencing and expression in Escherichia coli of the ptsI gene encoding enzyme I of the phosphoenolpyruvate:sugar phosphotransferase transport system from Streptococcus salivarius.

We present the cloning and sequencing of the ptsI gene, encoding enzyme I (EI) of the phosphoenolpyruvate (PEP): sugar phosphotransferase (PTS) transport system from Streptococcus salivarius. The ptsI gene corresponds to an open reading frame of 1731 nucleotides, which translates into a putative 577-amino acid (aa) protein with a M(r) of 62,948 and a pI of 4.49. The EI was produced in Escherichia coli under the control of its own promoter located immediately upstream of ptsI, a situation never previously reported for any other gene coding for an EI. The deduced aa sequence of the S. salivarius EI shows a high degree of similarity with the E. coli EI and the EI moiety of the multiphosphoryl transfer protein from Rhodobacter capsulatus. The S. salivarius EI also shares a highly conserved aa cluster with a non-PTS protein, the maize pyruvate:orthophosphate dikinase. The conserved cluster is located in a domain which is hypothesized to be the PEP-binding site.

Amino Acid Sequence↗
PubMed1992

Molecular analysis of isolates of Streptococcus suis capsular type 2 by restriction-endonuclease-digested DNA separated on SDS-PAGE and by hybridization with an rDNA probe.

This study was undertaken to assess the discriminatory value of restriction endonuclease (RE) digestion patterns of Streptococcus suis chromosomal DNA using polyacrylamide gel electrophoresis (SDS-PAGE) and DNA-rDNA hybridization. For the RE digestion patterns, DNAs were digested separately with the enzymes BamHI and BglII and the resultant fragments were separated by SDS-PAGE. An Escherichia coli rDNA probe derived from pKK3535 was used for the hybridization. Twenty-three S. suis capsular type 2 isolates recovered from diseased and clinically healthy pigs, from a human case, and from a cow were compared in this study. The majority of isolates associated with septicaemia belonged to one restriction endonuclease analysis (REA) profile group. Isolates associated with pneumonia belonged either to the REA profile group of isolates associated with septicaemia or to a second REA profile group. The REA profiles of isolates from clinically healthy animals were more heterogeneous. The REA profile of the type 2 reference strain, S735, which was originally isolated from a pig, was very different from those of the porcine and bovine isolates but similar to the profile of the human isolate. The profiles obtained after rDNA hybridization were more homogeneous. Although different patterns were detected in the 23 isolates, there was no correlation between the source of the isolate and the patterns observed with this technique.

Animals↗
PubMed1991

Construction, expression and release of hybrid colicins.

Colicins A and E1 are two pore-forming colicins sharing homology in their C-terminal domains but not in their N-terminal or central domains. Using site-directed mutagenesis, restriction sites were inserted at the proper locations to allow recombination of these domains. Six different constructs were obtained. All these proteins were expressed in Escherichia coli and properly recognized by monoclonal antibodies directed against epitopes located in different domains of colicin A. Out of the six hybrids, only two were released to the extracellular medium. Immunocytolocalization indicated that some of the hybrids aggregated within the cytoplasm. With some hybrids, the defect in release was related to a defect in synthesis of the lysis protein that normally promotes release.

Antibodies, Monoclonal↗
PubMed1991

Individual domains of colicins confer specificity in colicin uptake, in pore-properties and in immunity requirement.

Six different hybrid colicins were constructed by recombining various domains of the two pore-forming colicins A and E1. These hybrid colicins were purified and their properties were studied. All of them were active against sensitive cells, although to varying degrees. From the results, one can conclude that: (1) the binding site of OmpF is located in the N-terminal domain of colicin A; (2) the OmpF, TolB and TolR dependence for translocation is also located in this domain; (3) the TolC dependence for colicin E1 is located in the N-terminal domain of colicin E1; (4) the 183 N-terminal amino acid residues of colicin E1 are sufficient to promote E1AA uptake and thus probably colicin E1 uptake; (5) there is an interaction between the central domain and C-terminal domain of colicin A; (6) the individual functioning of different domains in various hybrids suggests that domain interactions can be reconstituted in hybrids that are fully active, whereas in others that are much less active, non-proper domain interactions may interfere with translocation; (7) there is a specific recognition of the C-terminal domains of colicin A and colicin E1 by their respective immunity proteins.

Bacterial Outer Membrane Proteins↗
PubMed1991

HPr polymorphism in oral streptococci is caused by the partial removal of the N-terminal methionine.

HPr is a small phosphocarrier protein of the bacterial phosphoenolpyruvate:sugar phosphotransferase system involved in the transport and phosphorylation of sugars. It has recently been reported that streptococci possess two forms of HPr having identical biochemical properties. In this communication, we show by N-terminal amino-acid sequencing and by ion-spray mass spectroscopy that these two forms differ by the presence or the absence of the N-terminal methionine.

Amino Acid Sequence↗
PubMed1989

Interactions of colicin A domains with phospholipid monolayers and liposomes: relevance to the mechanism of action.

The colicin A polypeptide chain (592 amino acid residues) contains three domains which are linearly organized and participate in the sequential steps involved in colicin action. We have compared the penetrating ability in phospholipid monolayers and the ability to promote vesicle fusion at acidic pH of colicin A and of protein derivatives containing various combinations of its domains. The NH2-terminal domain (171 amino acid residues), required for translocation across the outer membrane, has little affinity for dilauroylphosphatidylglycerol (DLPG) monolayers at all pHs tested. The central domain has a pH-dependent affinity, although lower than that of the entire colicin A. The COOH-terminal domain contains a high-affinity lipid binding site, but in addition an electrostatic interaction is required as a first step in the process of penetration into negatively charged DLPG films. In contrast to the constructs containing the ionophoric domain, the NH2-terminal domain alone has no fusogenic activity for liposomes. These results are discussed with regard to the mechanism of entry and action of colicin A in sensitive cells. Our results suggest the existence of a pH-dependent interaction between the receptor binding domain (amino acid residues 172-388) and the pore-forming domain of colicin A (amino acid residues 389-592).

Binding Sites↗
PubMed1989

Comparison of the uptake systems for the entry of various BtuB group colicins into Escherichia coli.

Colicins A, E1, E2 and E3 belong to the BtuB group of colicins. The NH2-terminal region of colicin A is required for translocation, and defects in this region cannot be overcome by osmotic shock of sensitive cells. In addition to BtuB, colicin A requires OmpF for efficient uptake by sensitive cells. The roles of BtuB and OmpF in translocation and binding to the receptor of the colicins A, E1, E2 and E3 were compared. The results suggest that for colicin A OmpF is used both as a receptor and for translocation across the outer membrane. In contrast, for colicin E1, OmpF is used neither as a receptor nor for translocation. For colicins E2 and E3, the situation is intermediate: only BtuB is used as a receptor but both BtuB and OmpF are involved in the translocation step.

Bacterial Outer Membrane Proteins↗
PubMed1988

Site of action of a gonococcal growth inhibitor produced by Staphylococcus haemolyticus.

The inhibitory substance produced by Staphylococcus haemolyticus strain no. 7 acts on growing as well as resting gonococcal cells, as shown by reductions in viable counts. The optical density of these cell suspensions was only slightly reduced. The inhibitor caused lysis of gonococcal spheroplasts at 24 degrees C and 37 degrees C, but was much less active at 4 degrees C. Acting on intact gonococcal cells, the inhibitor caused a temperature-dependent release of radioactive cytoplasmic material. Electronmicroscopy showed that treated suspensions contained ghost cells with the cell envelope relatively intact. Our results suggest that the inhibitor may act on the cytoplasmic membrane of the gonococcal cell causing cytoplasmic leakage and, eventually, death.

Antimicrobial Cationic Peptides↗
PubMed1988

Functional domains of colicin A.

A large number of mutations which introduce deletions in colicin A have been constructed. The partially deleted colicin A proteins were purified and their activity in vivo (on sensitive cells) and in vitro (in planar lipid bilayers) was assayed. The receptor-binding properties of each protein were also analysed. From these results, we suggest that the NH2-terminal region of colicin A (residues 1 to 172) is involved in the translocation step through the outer membrane. The central region of colicin A (residues 173 to 336) contains the receptor-binding domain. The COOH-terminal domain (residues 389 to 592) carries the pore-forming activity.

Binding Sites↗
PubMed1988

[Reintegration of people suffering from schizophrenia : marginalized for life ?].

This article develops a theory based on 10 years of involvement in an external psychiatric clinic identified as Mental Health Centre (<< Centre de santé mentale >>). Our first objective is to break the isolation in which our subjects have been living and to succeed in establishing significant ties with them. Our work concentrates on people suffering from schizophrenia that we meet daily in groups. In the article, we present a profile of this clientele, our conception of their way of adaptating to life, their therapeutic course and the framework in which it has evolved. Given the development of newly acquired skills as a result of these privileged ties, how do the beneficiaries transpose them into community life? While some succeed in using their new skills and others don't, the latter group must face the necessity of an adapted social reinsertion. This is the reason why we have decided to focus on the dynamic understanding of these people and the particular conditions of their social reinsertion.

English Abstract↗
PubMed1984

Chemical and biological characterization of a gonococcal growth inhibitor produced by Staphylococcus haemolyticus isolated from urogenital flora.

The purified antigonococcal substance produced by Staphylococcus haemolyticus no. 7 has shown a broad antigonococcal spectrum and a narrow antibacterial spectrum. The inhibitor produced in vitro was also active in the guinea pig subcutaneous chamber. The inhibitor has shown hemolytic activity; the human, horse, and mouse erythrocytes were the most susceptible. Hemolytic and antigonococcal activities were inhibited in the presence of phosphatidylcholine. The amino acid composition of the antigonococcal substance was characterized by the absence of proline, tyrosine, histidine, cysteine, and tryptophan. The molecular weight was found to be 2,565, and the major isoelectric points were 4.8 and 4.9 in the presence of 8 M urea and 4.6 without urea. The inhibitor has some properties similar to those of the delta toxin of Staphylococcus aureus, although the two substances are different based mainly on their chemical characteristics. Also an antiserum directed against the gonococcal inhibitor did not give a precipitation line with the delta toxin, indicating that the two substances are antigenically unrelated.

Amino Acids↗
PubMed1984

[Group therapy through graphic art.].

In this article we describe and comment upon two graphic art therapy groups. Is it possible to accomplish work of a psycho-therapeutic nature in such groups? We attempt to respond to this question using examples taken from the experience of these different groups. First, we explain how the groups operate, our method of intervention, the general evolution, and finally, we present, in an in-depth fashion, the progress of one of the participants.

English Abstract↗
PubMed1983

Comparison of the in vivo multiplication of 10 BCG substrains in three strains of mice: no correlation with their antitumour activity.

The dynamics of in vivo multiplication of 10 BCG substrains were studied in three different strains of mice: CF-1, B6D2F1/J and C57BL/6. For each substrain, the number of CFU recovered from the spleen of mice previously injected intravenously with BCG was determined weekly for 5 consecutive weeks. On the basis of the statistical analysis of the results, the BCG substrains were classified for each strain of mice in three groups of virulence: high, intermediate and low. The Russian and Japanese substrains were found to be the most virulent, and the Glaxo and Tice 946 BL (I2) the least virulent substrains. No correlation could be established between in vivo multiplication and antitumour activity of each BCG substrain as estimated in preventive treatment against three tumour systems: Ehrlich carcinoma in CF-1 mice, L1210 leukaemia in B6D2F1/J mice, and B16 melanoma in C57BL/6 mice.

Animals↗
PubMedDate not supplied

Uptake across the cell envelope and insertion into the inner membrane of ion channel-forming colicins in E coli.

Pore-forming colicins exert their lethal effect on E coli through formation of a voltage-dependent channel in the inner (cytoplasmic-membrane) thus destroying the energy potential of sensitive cells. Their mode of action appears to involve 3 steps: i) binding to a specific receptor located in the outer membrane; ii) translocation across this membrane; iii) insertion into the inner membrane. Colicin A has been used as a prototype of pore-forming colicins. In this review, the 3 functional domains of colicin A respectively involved in receptor binding, translocation and pore formation, are defined. The components of sensitive cells implicated in colicin uptake and their interactions with the various colicin A domains are described. The 3-dimensional structure of the pore-forming domain of colicin A has been determined recently. This structure suggests a model of insertion into the cytoplasmic membrane which is supported by model membrane studies. The role of the membrane potential in channel functioning is also discussed.

Amino Acid Sequence↗
PubMedDate not supplied

Stimulation of non-specific anti-tumour resistance in the mouse using cell wall preparations from four BCG substrains.

The comparative anti-tumour activity of cell walls, deproteinized cell walls and cell wall skeleton (CWS) isolated from four BCG substrains (Canadian, Russian, Glaxo and Swedish) was determined against Ehrlich ascitic carcinoma in CF-1 mice and against L1210 lymphoid leukaemia in B6D2F1/J mice. In Ehrlich carcinoma, the protocol of injection was shown to be of critical importance, since high and low levels of protection and even facilitation of tumour growth were observed according to the protocol used. The oil emulsion used as the vehicle for injection of the fractions (control groups) induced facilitation of tumour growth with some protocols. The highest levels of protection were observed when treatment was performed on day -6 for a single i. p. injection or on days -14 and 0 for two injections. Using the former protocol of immunization, the highest level of protection was detected with cell walls and deproteinized cell walls. CWS isolated from the Canadian and Glaxo substrains were found to be inactive, whereas those isolated from the Swedish and Russian substrains induced significant levels of protection. In L1210 leukaemia, very few preparations induced significant protection. CWS preparations did not induce protection. Canadian and Russian deproteinized cell walls and Canadian cell walls (100 and 250 micrograms, respectively) induced some protection. The use of different protocols of injection did not increase the anti-tumour activity of the Canadian cell walls. The most active preparation in both tumour systems was that of Canadian deproteinized cell walls.

Animals↗