Cellular transformation, tyrosine kinase oncogenes, and the cellular adhesion plaque

The study of adhesion plaques in normal and transformed cells provides a series of phenotypic markers by which the process of transformation can be followed. Several proteins which are concentrated in adhesion plaques have now been identified; a few of these can act as targets for tyrosine kinase. In an attempt to characterize the relationship between tyrosine phosphorylation and cell transformation, the reactions of three such proteins – vinculin, talin and integrin – with a range of tyrosine kinase oncogene products have been studied in detail.     

Effects of nitrous oxide on mitochondrial and cell respiration and growth in Distichlis spicata suspension cultures

The reversible inhibition of respiratory activity could provide a novel approach to the preservation of traditionally hard to store plant germplasm such as clonal materials and recalcitrant seed. The gaseous anesthetic nitrous oxide caused a reversible, dose-dependent, partial inhibition of dioxygen utilization in mitochondrial particles isolated from cell suspension cultures of the salt-tolerant marsh grass Distichlis spicata, with maximal inhibition of 33% after 30 minutes exposure to an atmosphere of 80% nitrous oxide plus 20% oxygen. Respiration of whole cells required longer time to be affected by the anesthetic, and was reversibly inhibited an average of 19% when measured using a differential respirometer. Exposure to 80% nitrous oxide plus 20% oxygen for up to 10 days caused no measurable effect on cell growth.Abbreviations PCV packed cell volume - EDTA sodium ethylenediaminetetraacetic acid - BSA bovine serum albumin - MOPS 3(N-morpholino) propanesulfonic acid - TMPD N,N,N',N'-tetramethyl-p-phenylene diamine - STP standard temperature and pressure     

Genes, bees and ecosystems: The evolution of a common interest among individuals

Adam Smith proposed that individuals can share sufficient interest in their society's welfare to profit by cooperating for its benefit and by jointly suppressing behavior hurtful to it. Similarly, a genome's genes share a common interest in 'honest' meiosis, which ensures that alleles can spread only by benefitting their carriers. Honeybee workers express a common interest in raising their queen's, rather than their half-sisters', eggs by eating eggs laid by half sisters. Can analogous principles explain the evolution of harmony at other levels of biological organization, such as ecosystems or organismic development?     

Observation of manganese(II)-ligand superhyperfine couplings in complexes with proteins by electron spin-echo spectroscopy

Pulsed electron paramagnetic resonance spectroscopy has been used to detect Mn(II)-ligand superhyperfine couplings in complexes with creatine kinase and in the Mn(II) metalloprotein concanavalin A. Electron spin-echo envelopes from Mn(II), bound in these complexes, are modulated by superhyperfine interactions between Mn(II) and nearby, weakly coupled nuclear spins. The characteristic frequencies of the modulations were obtained by Fourier transformation of the three-pulse, spin-echo envelopes. In transition-state analogue complexes of creatine kinase (enzyme-MnIIADP-anion-creatine), superhyperfine interactions from the directly coordinated nitrogen of the thiocyanate ligand give envelope modulations. The source of the modulations was confirmed by measurements with the 14N and 15N forms of thiocyanate. On the other hand, the nitrogen of coordinated nitrate, which is two bonds removed from the paramagnetic center, does not produce detectable modulations. In spectra for Mn(II) concanavalin A, envelope modulations are detected due to the nitrogen of the coordinated histidine residue. Complexes prepared in 2H2O give strong signals due to weakly coupled 2H. For Mn(II)-doped single crystals of sodium pyrophosphate, signals are observed in the frequency domain spectra that are due to coupling from 31P. Phosphorus signals from the ADP ligand in complexes with creatine kinase show approximately the same coupling constant but have a much broader line width.     

Interaction of tumour promoters with epithelial cells in culture : An immunofluorescence study

12-O-tetradecanoyl phorbol-13-acetate (TPA) has a profound and rapid influence on the cytoskeleton of Madin-Darby Canine Kidney (MDCK) cells. Within 10 min, TPA induces a rapid change in morphology, from a flat, cuboidal state to a rounded or elongated morphology in which the cell membranes become convoluted. Concomitant with this morphological change is a rapid dissolution of stress fibres and a redistribution of F-actin from microfilament bundles to a membrane or sub-membranous location. The rearrangement of actin is paralleled by a rearrangement of alpha-actinin and a reduction in the number of vinculin-containing adhesion plaques. Unusual F-actin configurations are often found emanating from a perinuclear location, usually containing alpha-actinin and terminating in a vinculin-containing adhesion plaque. The cytoskeletal rearrangements occur in the presence of inhibitors of protein synthesis or oxidative phosphorylation, but do not occur if glycolysis is also inhibited. The rearrangements are partly abrogated by the presence of cytochalasin B (CB). Despite these dramatic changes in microfilaments the polymerization state of actin remained unaltered after TPA treatment. Furthermore, although changes in the movement of membrane lipids have been reported, no obvious differences in the ability of glycoproteins to redistribute in the plane of the membrane were found as judged by FITC-concanavalin A (conA) induced patching. The rapidity of the morphological response of MDCK cells to TPA indicates that the cytoskeleton is one of the primary targets of TPA, but that tumour promoters differ from RNA tumour viruses in their effect on the state of actin polymerization.