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T David-Pfeuty

Publications and source records attributed to T David-Pfeuty.

26 records · Page 2Linked to original sources
PubMed1983

The effect of microtubule disruption on the distribution of the cytoskeletal proteins, vinculin, alpha-actinin, and actin in normal and RSV-transformed fibroblasts.

By double indirect immunofluorescence and interference electron microscopy, we have observed the effect of microtubule disruption by antimitotic drugs and coldness treatment on the distribution of adhesion sites and of the three cytoskeletal proteins, vinculin, alpha-actinin, and actin in normal rat cells and in rat cells transformed by Rous sarcoma virus. This study shows that the state of organization of the microtubule--intermediate filament complex modulates the location and the arrangement of intracellular structures containing vinculin, alpha-actinin, and actin in normal as well as in transformed cells. The most important alterations are observed in transformed cells on the distribution of the rosette clusters that have been shown to characterize the transformation by Rous sarcoma virus [8]. These results suggest the microtubule-intermediate filament complex is directly or indirectly connected with the microfilament network.

Actinin↗
PubMed1980

Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus.

It was recently shown by combined immunofluorescence and interference reflection microscopy that a protein named vinculin, along with alpha-actinin, is concentrated at focal adhesion plaques inside cultured normal fibroblasts [Geiger, B. (1979) Cell 18, 193-205]. These plaques are the discrete, isolated sites of strong adhesions formed between the ventral surfaces of the cells and the substrata on which they are grown. We show that after transformation of fibroblasts by Rous sarcoma virus a majority of the cells have many fewer focal adhesion plaques and now exhibit a cluster of small patches that are immunolabelled for both vinculin and alpha-actinin. Such a cluster (rosette) is located near the ventral surface of the cell, usually partly under the nucleus. The significance that these altered distributions of vinculin and alpha-actinin may have for the rounding up and loss of adherence of transformed cells is discussed.

Actinin↗
PubMed1979

Effect of antimitotic drugs on tubulin GTPase activity and self-assembly.

Microtubule inhibitors can be classified into two categories: 1) those which inhibit the polymerization-dependent GTPase activity of phosphocellulose-purified tubulin, but induce a significant polymerization-independent GTPase activity (e.g. colchicine, griseofulvine, daunorubicine); 2) those which inhibit the GTPase activity associated with tubulin polymerization and that induced by inhibitors of the first class (e.g. the vincaalkaloids and podophyllotoxin). The colchicine-stimulated GTPase activity of tubulin appears to be due to the tubulin.colchicine complex. This suggests that colchicine inhibits tubulin assembly by binding to a tubulin-tubulin interaction site required for the polymerization-dependent GTPase activity and induces by itself a tubulin conformational change that leads to polymerization-independent GTPase activity. Stoichiometry of inhibition by vinblastine of the colchicine-stimulated GTPase activity is 1:2. On the other hand, the inhibition by vinblastine of the tubulin self-assembly and of the polymerization-dependent GTPase activity is strongly substoichiometric at the beginning of the polymerization reaction, 1 vinblastine molecule inhibiting the ability of 10 tubulin dimers to polymerize and to hydrolyze the GTP. However, at the polymerization plateau, the inhibition effect by vinblastine appears to be lower, suggesting a selective action of vinblastine on the early stages of the polymerization reaction.

Animals↗
PubMed1977

Guanosinetriphosphatase activity of tubulin associated with microtubule assembly.

Tubulin, purified by cycles of assembly followed by phosphocellulose chromatography, exhibits a characteristic GTPase activity that is polymerization dependent and can be attributed to the tubulin itself. This activity has been observed, in a standard reassembly buffer containing low Mg2+, under three conditions that induce microtubule assembly: in the presence of microtubule-associated proteins, in the presence of DEAE-dextran, or after addition of high Mg2+ and glycerol. The phosphocellulose-purified tubulin showed no GTPase activity under the following nonpolymerizing conditions: in buffer with low Mg2+ in the absence of microtubule-associated proteins or DEAE-dextran, in buffer with high Mg2+ and glycerol at tubulin concentrations below the critical concentration, or when microtubule assembly was inhibited by vinblastine. Colchicine, on the other hand, while blocking microtubule assembly, induced a significant GTPase activity in the phosphocellulose-purified tubulin. During the process of assembly, GTP appears to be hydrolyzed as a free tubulin dimer polymerizes into a microtubule. A constant GTPase activity when polymerization equilibrium is reached apparently reflects the cyclic polymerization-depolymerization of tubulin dimers at the ends of the microtubules.

Animals↗
PubMed1975

Regulation of binding properties of the nicotinic receptor protein by cholinergic ligands in membrane fragments from Torpedo marmorata.

Exposure of receptor-rich membrane fragments from Torpedo marmorata to carbamylcholine causes a slow (half-time of 5--10 min) and reversible change of properties of the cholinergic receptor protein manifested by a decrease of the initial rate of Naja nigricollis alpha-[3H]toxin binding in the presence of carbamylcholine. This change corresponds to a 5- to 20-fold increase of affinity for carbamylcholine. Other agonists, acetylcholine, phenyltrimethylammonium, show the same effect but not the antagonists d-tubocurarine and flaxedil. Decamethonium and hexamethonium show little, if any, agonistic effect in vitro on the same membrane fragments but cause the affinity change. This regulatory property can be lost after aging of the preparation of membrane fragments. Since the affinity increase progresses with a similar time course as the decrease of amplitude of the permeability response consecutive to agonist preincubation, it is proposed that, in the membrane at rest, the receptor protein is present under a state of low affinity for agonists and that the reversible stabilization by the agonists of a high affinity state corresponds to the "pharmacological desensitization" of the system as predicted by one of the models of Katz and Thesleff.

Acetylcholine↗