Construction of hybrid plasmids containing the lysA gene of Escherichia coli: studies of expression in Escherichia coli and Saccharomyces cerevisiae

Summary The lysA gene of Escherichia coli has been cloned from a transducing phage on various plasmids, present in different copy numbers in bacterial cells. Synthesis of the product of this gene, diaminopimelate (DAP)-decarboxylase, and its regulation have been studied. Expression does not follow a simple gene dosage effect, maximal expression already being obtained with a six-copy plasmid. This result suggests that either a positive or an autogenous regulatory mechanism is involved. We also used one of the hybrid plasmids to look for expression of the bacterial lysA gene in Saccharomyces cerevisiae. The results indicate that the product of the E. coli gene is not actively translated in yeast.     

The relA locus and the regulation of lysine biosynthesis in Escherichia coli

Summary The allelic state of relA influences the phenotype of Escherichia coli strains carrying the lysA22 mutation: lysA22 relA strains are Lys^– where lysA22 relA ⁺ strains grow (slowly) in the absence of lysine. This physiological effect has been related to an effect of the expression of the relA locus on the regulation of lysine biosynthesis. The fully derepressed levels of some lysine enzymes (aspartokinase III, aspartic semialdehyde dehydrogenase, dihydrodipicolinate reductase) are observed under lysine limitation only in rel ⁺ strains. And the induction of DAP-decarboxylase by DAP is much higher in rel ⁺ than in rel ^– strains when an amino acid limitation of growth is also realised. These results are in agreement with the hypothesis of Stephens et al. (1975) on a possible role of the stringent regulation as a general signal for amino acid deficiency.     

Iron-porphyrin catalysis of the oxidative dealkylation of para-nitro-anisole and 7-ethoxycoumarin by cumylhydroperoxide: a possible model for the corresponding cytochrome P 450-dependent reactions

A biphasic system containing an iron porphyrin, Fe (TPP) (C1)¹ or [Fe(TPP)]₂O, efficiently catalyzes the cumyl-or tertiobutyl-hydroperoxide-supported dealkylation of p-nitroanisole and 7-ethoxycoumarin to the corresponding phenol and formaldehyde. Stoichiometric amounts of iron porphyrin and hydroperoxide give a quantitative reaction. Catalytic amounts of iron porphyrin give reaction rates and yields which are proportional to substrate concentration. With increasing hydroperoxide concentrations, the rates level offto limit values and the yield rapidly decreases. The maximum rates obtained approach those of the reactions mediated by cytochrome P 450-dependent monooxygenases.     

Réflexions critiques sur l'écologieet la systématique des Lingules actuelles et fossiles

New topics on ecology and systematics of recent and fossil Lingulids lead to an obvious revision of our knowledges on this zoological group. At first, the recent species need systematics and taxonomy on the bases of new described specific criteria (as, morphology of deltidial areas, muscle disposition); the results are briefly indicated. But, in fossil species, disorder and disparity of used characteristics are emphasized.The general conceiving on ecology of Lingulids are reviewed and discussed, especially on bathymetry and salinity; sediment and oxygenation conditions; taphocoenosis and lingulid «communities. On recent species, all these points are also studied, especially some ecological requirements (salinity, bathymetry, grain size), and mechanism of burrowing ability, burrow living positions in the sediments, as shell preservations after death and fossilization, to facilitate the paleobiotope interpretations. Recent animals are euryhalin, surviving at salinities from about 13 to 42‰; they could be considered as well adapted to waters with strong salinity fluctuations. They show preference to fine sand bottoms (lowest particle size about 40–60 μm). Their bathymetric distribution occurs between 0 and about 500 m (Lingula especially between 5–50 m; Glottidia 15–70 m). The isotherms 8–10°C seem to restrict their geographical and bathymetric distribution.Therefore, some post-palaeozoic lingulid bedsare studied or redescribed on the bases of the above discussed characteristics, and new interpretations on the environmental situation are given (Trias of Vosges Mountains; Oligocene from Japan; Eocene of London Basin). More caution must be used in study of fossil Lingulids that are not especially animals living in infralittoral bottoms with low salinity and deficient oxygenation, as generally accepted.     

Mitochondrial protein alteration in active brown fat: A sodium dodecyl sulfate-polyacrylamide gel electrophoretic study

The polypeptide composition of mitochondria isolated from the brown adipose tissue of control rats (bred at 22°C) or cold-exposed animals (bred at 6°C) was compared using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A striking increase in the content of an unknown polypeptide (apparent molecular weight 32,000 daltons) was found after cold adaptation, a phenomenon which was reversed during re-adaptation to a normal temperature. This protein seems to be localized in the membrane fraction of the mitochondria.     

Reciprocal influence of connexins and apical junction proteins on their expressions and functions

Membranes of adjacent cells form intercellular junctional complexes to mechanically anchor neighbour cells (anchoring junctions), to seal the paracellular space and to prevent diffusion of integral proteins within the plasma membrane (tight junctions) and to allow cell-to-cell diffusion of small ions and molecules (gap junctions). These different types of specialised plasma membrane microdomains, sharing common adaptor molecules, particularly zonula occludens proteins, frequently present intermingled relationships where the different proteins co-assemble into macromolecular complexes and their expressions are co-ordinately regulated. Proteins forming gap junction channels (connexins, particularly) and proteins fulfilling cell attachment or forming tight junction strands mutually influence expression and functions of one another.     

Spatial structure of a subtidal macrobenthic community in the Bay of Veys (western Bay of Seine, English Channel)

The spatial distribution of the muddy fine sand community from the Bay of Veys (western English Channel) were investigated during spring and autumn 1997. A grid of 55 and 54 sites was sampled in March and October, respectively, using two replicates per site of a Hamon grab (0.25 m²) for macrofauna collection and an additional one for sediment analysis. A total of 172 species were sampled with a dominance of polychaetes, followed by crustaceans and bivalves. The species richness and abundance show low temporal changes despite higher values in October than in March. In March, the mean abundance was 165 ind. 0.5 m⁻²; in October, the mean abundance was 212 ind. 0.5 m⁻². Four assemblages from the Abra alba-Pectinaria koreni community were identified corresponding to a bathymetric and sedimentary gradient from muddy fine sands with high levels of fine particles in shallow water to fine sands in deeper water. The discussion focuses on factors prevailing on the spatial structure of sandy communities in the English Channel.     

A physiological and molecular study of the effects of nickel deficiency and phenylphosphorodiamidate (PPD) application on urea metabolism in oilseed rape (Brassica napus L.)

Background and aims

Urea is the major nitrogen (N) form supplied as fertilizer in agriculture. However, urease, a nickel-dependent enzyme, allows plants to use external or internally generated urea as a nitrogen source. Since a urease inhibitor is frequently applied in conjunction with urea fertilizer, the N-metabolism of plants may be affected. The aim of this study was to determine physiological and molecular effects of nickel deficiency and a urease inhibitor on urea uptake and assimilation in oilseed rape.

Methods

Plants were grown on hydroponic solution with urea as the sole N source under three treatments: plants treated with nickel (+Ni) as a control, without nickel (?Ni) and with nickel and phenylphosphorodiamidate (+Ni+PPD). Urea transport and assimilation were investigated.

Results

The results show that Ni-deficiency or PPD supply led to reduced growth and reduced ¹⁵N-uptake from urea. This effect was more pronounced in PPD-treated plants, which accumulated high amounts of urea and ammonium. Thus, Ni-deficiency or addition of PPD, limit the availability of N and decreased shoot and root amino acid content. The up-regulation of BnDUR3 in roots indicated that this gene is a component of the stress response to nitrogen-deficiency. A general decline of glutamine synthetase (GS) activity and activation of glutamate dehydrogenase (GDH) and increases in its expression level were observed in control plants. At the same time, in (?N) or (+Ni+PPD) treated plants, no increases in GS or GDH activities and expression level were found.

Conclusions

Overall results showed that plants require Ni as a nutrient (while most widely used nutrient solutions are devoid of Ni), whether they are grown with or without a urea supply, and that urease inhibitors may have deleterious effects at least in hydroponic grown oilseed rape.