Management of time by the cell and by the organism

Time-dependent regulations of cells and organisms can be analysed at different levels. One of these levels is the periodicity of cell functions such as cell division, metabolic processes (generation of ATP by glycolysis or oxidative mitochondrial processes) and the biosynthesis of cell constituents. Studies carried out on unicellular eukaryotes revealed the periodic, oscillatory nature of most of these processes. Time constants of these reactions vary from nanoseconds to hours-days, necessitating coupling mechanisms. Comparative studies revealed the coupling of the rapid processes (mitochondrial ATP generation) to the slower rhythms of the biosynthetic processes of macromolecules. Adenine nucleotides are involved in the coupling mechanisms between rapid and slow processes ("the slow dance of life to the music of time"). The mechanisms underlying these rhythmic processes involve either key allosteric regulatory enzymes (PFK for glycolysis) or "desensitization" of receptors by phosphorylation-dephosphorylation. At the organismic level the study of rhythmic processes is illustrated by the periodicity of heart beats, shown to exhibit multifractality, following apparently the formalism of deterministic chaos. Another example is the rhythmic oscillatory discharges of neuronal networks. The existence of subrhythmes mostly of epigenetic nature, facilitated probably the progressive adjustment of cells during evolution to the slow increase of day time since the separation of the moon from the earth. We analysed the mechanisms underlying the decline of these processes during aging. Loss of receptors or/and their uncoupling from their transmission pathway appear to be involved in most of these processes of decline. One conclusion of this review is the importance of epigenetic mechanisms both in the genesis and in the decline of these rythmic processes involved in time keeping by the cell.     

Amelioration of the teratogenicity of cadmium by the metallothionein induced by bismuth nitrate

The participation of maternal hepatic metallothionein (MT) in the amelioration of cadmium teratogenicity in mice was examined. Pretreatment with bismuth nitrate (subcutaneously) ameliorated the teratogenicity, including exencephaly and abnormalities of the axial skeleton, caused by a single intraperitoneal injection of cadmium sulfate. Pretreatment with bismuth nitrate for 3 days induced MT drastically in maternal liver and kidney. Six and 24 hr after the injection of cadmium sulfate, the accumulation of maternal hepatic cadmium increased and that in the decidua, including embryos, decreased after pretreatment with bismuth nitrate. Mouse embryos on day 7 of gestation were cultured for 48 hr. Exposure to cadmium sulfate in vitro induced unfused brain fold, which corresponds to exencephaly in vivo. From the in vitro experiment, it was suggested that the teratogenicity of cadmium on day 7 of gestation is a direct action against the mouse embryo. In the present experiment it was suggested that pretreatment with bismuth nitrate induced maternal hepatic and renal MT; cadmium was therefore trapped and detoxicated, and consequently embryos were exposed to a lower concentration of cadmium.     

Inhibition by diphenols of the induction of micronuclei by gamma-radiation

A study was made of the influence of diphenols (for instance, resorcinol, hydroquinone, and pyrocatechin) on gamma-radiation induction of micronuclei (1.5 Gy). The position of the diphenol molecule hydroxyl group (the isomeric effect) was shown to influence their antimutagenic activity. This antimutagenic effect of the diphenols is associated with their ability to produce semiquinone and quinone forms which are peculiar for the process of oxidation of pyrocatechin (ortho-) and hydroquinone (para-) as opposed to resorcinol (meta-position of the hydroxyl group).     

To live or die by the sword: the regulation of apoptosis by the proteasome

The proteasome has been implicated in the control of apoptosis by modulating the levels of both pro- and antiapoptotic molecules. A recent study published in the April 9th issue of Molecular Cell reveals that caspase-dependent inactivation of the proteasome can amplify the activation of apoptosis.     

Interference by Tris buffer in the estimation of protein by the Lowry procedure

Tris buffers were found to distort the measurement of protein by the Lowry method both by decreasing chromophore development with protein and by contributing blank color. Tris at an assay concentration of 0.37 mm markedly affects measured results. Similar Tris effects were observed at all wavelengths between 450 and 800 nm and with diverse protein samples. The distortion due to Tris is not correctable by simple blank correction, but it can be overcome by incorporating the same amount of Tris in the standards used. The distortion at Tris concentrations <0.15 mm appears to be within tolerable limits. No interference or distortion was observed with sodium phosphate buffer to an assay concentration of 40 mm. An automated Lowry procedure is also presented which gives excellent correlation with the manual method and an average coefficient of variation of <4%.     

Structure of the oligomers obtained by enzymatic hydrolysis of the glucomannan produced by the plant Amorphophallus konjac

Dimers and trimers obtained by enzymatic hydrolysis of the glucomannan produced by the plant Amorphophallus konjac were analysed in order to obtain information on the saccharidic sequences present in the polymer. The polysaccharide was digested with cellulase and beta-mannanase and the oligomers produced were isolated by means of size-exclusion chromatography. They were structurally characterised using electrospray mass spectrometry, capillary electrophoresis, and NMR. The investigation revealed that many possible sequences were present in the polymer backbone suggesting a Bernoulli-type chain.     

Inactivation of isocitrate dehydrogenase by phosphorylation is mediated by the negative charge of the phosphate

The control of isocitrate dehydrogenase through phosphorylation is necessary for growth of Escherichia coli on acetate as the sole carbon source. To understand the mechanism by which phosphorylation inactivates isocitrate dehydrogenase, the sequence of icd, the isocitrate dehydrogenase structural gene, was determined and this information was used to construct mutants at the site of phosphorylation. Introduction of a negatively charged aspartate for the serine that is phosphorylated completely inactivates isocitrate dehydrogenase. Substitution of the serine with other amino acids results in a partially active enzyme in which both maximal velocity and interaction with substrates has been altered. Neither threonine nor tyrosine, when substituted for the serine at the phosphorylation site, is detectably phosphorylated by isocitrate dehydrogenase kinase.     

Alteration by mutation of the control by oxygen of the nar operon in Escherichia coli

Summary A nar-lac operon fusion was used to isolate a mutant in which the expression of the nar operon was no longer repressed by oxygen. The nar ^d mutation, located upstream of the nar structural genes, was found to be cis dominant; it led to independence from the Fnr protein which, in the wild-type strain, exerts a strict positive control on the nar operon. Both other known controls, nitrate induction and autoregulation, were unaffected. It is proposed that molecular oxygen controls the expression of nar via Fnr and that the nar ^d mutation affects the Fnr binding site of the narGHI control region.     

Taming the tiger by the tail: modulation of DNA damage responses by telomeres

Telomeres are by definition stable and inert chromosome ends, whereas internal chromosome breaks are potent stimulators of the DNA damage response (DDR). Telomeres do not, as might be expected, exclude DDR proteins from chromosome ends but instead engage with many DDR proteins. However, the most powerful DDRs, those that might induce chromosome fusion or cell‐cycle arrest, are inhibited at telomeres. In budding yeast, many DDR proteins that accumulate most rapidly at double strand breaks (DSBs), have important functions in physiological telomere maintenance, whereas DDR proteins that arrive later tend to have less important functions. Considerable diversity in telomere structure has evolved in different organisms and, perhaps reflecting this diversity, different DDR proteins seem to have distinct roles in telomere physiology in different organisms. Drawing principally on studies in simple model organisms such as budding yeast, in which many fundamental aspects of the DDR and telomere biology have been established; current views on how telomeres harness aspects of DDR pathways to maintain telomere stability and permit cell‐cycle division are discussed.