The effect of Ca2+ ions on DNA compaction in the complex with non-histone chromosomal protein HMGB1

The analysis of absorption and circular dichroism spectra in UV and IR regions showed that Ca2+ ions interact both with the phosphate groups of DNA and with the HMGB1 protein. Not only negatively charged C-terminal part of the protein molecule participates in interaction with metal ions but also its DNA-binding domains. The latter fact leads to the change of the mode of protein-DNA interaction. The presence of Ca2+ ions prevents formation of ordered supramolecular structures, specific for the HMGB1-DNA complexes, though promotes intermolecular aggregation. The structure of the complexes between DNA and the protein HMGB1 lacking C-terminal tail appears to be the most sensitive to the presence of Ca2+ ions. The data obtained allow to conclude that Ca2+ ions do not play a structural role in the HMGB1/DNA complexes and the presence of these ions is not necessary to DNA compaction in such systems.     

Control of Saccharomyces cerevisiae catalase T gene (CTT1) expression by nutrient supply via the RAS-cyclic AMP pathway.

In Saccharomyces cerevisiae, lack of nutrients triggers a pleiotropic response characterized by accumulation of storage carbohydrates, early G1 arrest, and sporulation of a/alpha diploids. This response is thought to be mediated by RAS proteins, adenylate cyclase, and cyclic AMP (cAMP)-dependent protein kinases. This study shows that expression of the S. cerevisiae gene coding for a cytoplasmic catalase T (CTT1) is controlled by this pathway: it is regulated by the availability of nutrients. Lack of a nitrogen, sulfur, or phosphorus source causes a high-level expression of the gene. Studies with strains with mutations in the RAS-cAMP pathway and supplementation of a rca1 mutant with cAMP show that CTT1 expression is under negative control by a cAMP-dependent protein kinase and that nutrient control of CTT1 gene expression is mediated by this pathway. Strains containing a CTT1-Escherichia coli lacZ fusion gene have been used to isolate mutants with mutations in the pathway. Mutants characterized in this investigation fall into five complementation groups. Both cdc25 and ras2 alleles were identified among these mutants.     

Contact-dependent regulation of growth of diploid human fibroblasts is dependent upon the presence of terminal galactose residues on plasma membrane glycoproteins

The growth of diploid human fibroblasts has previously been shown to be regulated mainly by the extent of cell-cell contacts [R. J. Wieser and F. Oesch (1986) J. Cell Biol. 103, 361], these contacts being effective only when terminal, beta-glycosidically linked galactose residues were present on plasma membrane glycoproteins. These studies, in which a high cell density in sparse cell cultures has been mimicked by the addition of immobilized plasma membrane glycoproteins, have been further extended to investigate the role of terminal galactose residues directly in cell cultures. The studies presented herein show that (i) culturing human fibroblasts in the presence of beta-galactosidase resulted in an approximately twofold higher saturation density, as well as a twofold higher proliferation rate at high cell densities when compared to the rates found in control cultures. (ii) The presence of alpha-lactalbumin in the culture medium, which acts as a modifier of the activity of galactosyltransferase, had the same effect as beta-galactosidase. (iii) Addition of the lectin I from Bandeiraea simplicifolia (BS I), which is specific for terminal galactose residues, resulted in an increase in the proliferation rate of cell cultures at high cell densities, while the proliferation was not affected at low cell densities. These data show that the presence of terminal, beta-glycosidically linked galactose is vital for the efficient growth control of normal cells.     

Low production “efficiency” of homoeotherm populations: a misunderstanding

Summary The production/respiration (P/R) ratios of homoeotherm populations are about one order of magnitude lower than those of poikilotherm populations. This has suggested to ecologists that homoeotherms are much less efficient converters of food energy into growth and reproduction than poikilotherms, the reason being the high cost of thermal homoeostasis. That this conclusion is wrong can be demonstrated by plotting three measures of production: individual growth rate; maximum rate of population increase; and natural rate of population increase, against rate of basal netabolism. It follows that to call the P/R ratio of communities a measure of production efficiency leads to confusion. If the P/R ratios of homoeotherm poulations are lower than those of poikilotherm populations it is not because their individual members are less efficient but because the low rate of natural increase happens to have been the most stable evolutionary solution for the organization of life cycle schedules in these animals.     

Purification and Characterization of Gallic Acid Decarboxylase from Pantoea agglomerans T71

Oxygen-sensitive gallic acid decarboxylase from Pantoea (formerly Enterobacter) agglomerans T71 was purified from a cell extract after stabilization by reducing agents. This enzyme has a molecular mass of approximately 320 kDa and consists of six identical subunits. It is highly specific for gallic acid. Gallic acid decarboxylase is unique among similar decarboxylases in that it requires iron as a cofactor, as shown by plasma emission spectroscopy (which revealed an iron content of 0.8 mol per mol of enzyme subunit), spectrophotometric analysis (absorption shoulders at 398 and 472 nm), and inhibition of the enzyme activity by 2,2′-bipyridyl, o-phenanthroline, and EDTA. Another interesting feature of this strain is the fact that it contains a tannase, which is used together with the gallic acid decarboxylase in a two-enzyme resting cell bioconversion to synthesize valuable pyrogallol from readily available tannic acid.     

The Aspergillus FlbA RGS domain protein antagonizes G protein signaling to block proliferation and allow development.

flbA encodes an Aspergillus nidulans RGS (regulator of G protein signaling) domain protein that is required for control of mycelial proliferation and activation of asexual sporulation. We identified a dominant mutation in a second gene, fadA, that resulted in a very similar phenotype to flbA loss-of-function mutants. Analysis of fadA showed that it encodes the alpha-subunit of a heterotrimeric G protein, and the dominant phenotype resulted from conversion of glycine 42 to arginine (fadA(G42R)). This mutation is predicted to result in a loss of intrinsic GTPase activity leading to constitutive signaling, indicating that activation of this pathway leads to proliferation and blocks sporulation. By contrast, a fadA deletion and a fadA dominant-interfering mutation (fadA(G203R)) resulted in reduced growth without impairing sporulation. In fact, the fadA(G203R) mutant was a hyperactive asexual sporulator and produced elaborate sporulation structures, called conidiophores, under environmental conditions that blocked wild-type sporulation. Both the fadA(G203R) and the fadA deletion mutations suppressed the flbA mutant phenotype as predicted if the primary role of FlbA in sporulation is in blocking activation of FadA signaling. Because overexpression of flbA could not suppress the fadA(G42R) mutant phenotype, we propose that FlbA's role in modulating the FadA proliferation signal is dependent upon the intrinsic GTPase activity of wild-type FadA.     

Metabolic rate and cost of growth in juvenile pike (Esox lucius L.) and perch (Perca fluviatilis L.): the use of energy budgets as indicators of environmental change

Summary Energy budgets of juvenile pike and perch (weighing approximately 3 g) were determined in experiments lasting up to 4 days, by simultaneously measuring oxygen consumption, food consumption, and growth of individual fish. Although the basic pattern of energy allocation was identical in the two species, perch subjected to constant light (PEL) showed faster growth, higher assimilation and conversion efficiency, and higher oxygen consumption than perch subjected to a short daylength regime (PED). The efficiency with which food energy was converted into body mass was 39±5% in PED but 49±4% in PEL. However, the “work coefficient” (increment of body mass/post-prandial increase of oxygen consumption: mg · μmol O _inf2 ^sup−1 ) differed only insignificantly between the two groups of perch, indicating that the metabolic cost of growth was unaffected by the manipulation of experimental conditions. This identifies the higher assimilation efficiency, i.e. the increased flow of food energy into the tissues as being the cause of accelerated growth of perch under the constant-light regime. In both species the maximum feeding-induced metabolic rate was 4 times higher than the lowest preprandial rate. In perch (which were kept on low rations before the experiments) the post-prandial metabolic rate increased steadily from day to day during the 4-day experiments, so that on the last day the rate of oxygen consumption exceeded the rate on the first day by about 41%. This investigation provides further evidence that the allocation of metabolic energy in fish is based on a flexible strategy which responds sensitively to changes in both internal and external conditions.     

Investigations on the ecophysiology of Geleia nigriceps Kahl (Ciliophora,Gymnostomata) inhabiting a sandy beach in Bermuda

Summary The dominant ciliate species Geleia nigriceps inhabiting a sheltered beach in Bermuda was characterized by its horizontal and vertical distribution on the beach in correlation with sediment composition and grain size, temperature, pH, oxygen, redox potential, and water coverage, its tolerance of high and low temperature under normoxic and oligoxic conditions, and its tolerance of extreme pH-values.From tolerance experiments it can be concluded that G. nigriceps is able to tolerate relatively high temperatures (up to 37.3°C) for only short time (e.g., when low tide occurs around noon on hot days).This species, inhabiting both the oxidized and the reduced milieu below the redox potential discontinuity layer of the beach, is only to a small degree dependent upon oxygen availability and can be regarded as a more or less habitual dweller of anoxic or oligoxic sediments. The functions relating LT₅₀ and exposure time were almost identical for normoxic and oligoxic conditions.Tolerance experiments revealed that pH values from 8.2 to 9.6 present no obstacle to G. nigriceps in its occupation of the upper millimeters of the sediment. In addition, it could be demonstrated that pO₂ has no effect on the tolerance of alkaline conditions.The vertical pattern of distribution of G. nigriceps correlated with water coverage during one period of outgoing tide suggests vertical migration. The upper sediment layer is more densely colonized than the lower ones at stations where water coverage exceeds 45%. A highly significant correlation exists between the population density of the upper centimeter and water coverage.Contribution No. 724 from the Bermuda Biological Station for Research     

The effects of season and temperature on D-lactate dehydrogenase, pyruvate kinase and arginine kinase in the foot of Helix pomatia L.

The effects of pH, season, environmental and experimental temperatures on the activities and kinetic parameters of D-lactate dehydrogenase, pyruvate kinase and arginine kinase from the foot of the pulmonate snail Helix pomatia were analyzed. Both in phosphate and Tris buffers D-lactate dehydrogenase was the enzyme with the most acid maximum, arginine kinase that with the most alkaline, whilst pyruvate kinase occupied an intermediate position. Pyruvate kinase activity, measured at 20 degrees C, was positively correlated with the environmental temperature at the moment of collecting the animal, whereas neither arginine kinase nor D-lactate dehydrogenase showed such a relationship. A seasonal study based on approximately 100 specimens established that arginine kinase activity remained the same throughout the year. Pyruvate kinase activity was slightly lower, and D-lactate dehydrogenase activity significantly higher, in winter than in summer animals. Snails subjected in spring to a short warm-up period before enzyme extraction showed extreme variability and some extraordinarily high values of pyruvate kinase activity, suggesting that either season or elevated temperature may have an immediate effect on the activity of this enzyme. Individual variability of all three enzymes ranges from 300 to 400%. The activities of pyruvate kinase and D-lactate dehydrogenase are strongly correlated in summer, forming a "constant-proportion-group", whereas in winter, with D-lactate dehydrogenase activity increasing and pyruvate kinase activity decreasing these two enzymes become "uncoupled". The Km value of pyruvate kinase is independent of experimental temperature between 10 and 25 degrees C, whereas that of D-lactate dehydrogenase and arginine kinase increases about three-fold within this range. Thus the temperature relationship of a single enzymic reaction cannot be used as an arguemnt for or against the occurrence of temperature compensation of whole animal metabolism. The possibility of modulation of enzyme activity by environmental temperature is discussed.