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A Ognibene

Publications and source records attributed to A Ognibene.

61 records · Page 4Linked to original sources

Changes in nuclear inositol phospholipids induced in intact cells by insulin-like growth factor I.

Swiss 3T3 cells were labelled for 36 hours with high levels of 3H-myo-inositol and the radioactivity in nuclear inositol phospholipids was measured. Treatment of cells for 2 minutes, but not for 4 hours, with mitogenic concentrations of insulin-like growth factor I and bombesin caused a slight decrease in PtdIns and more marked decreases in PtdInsP and PtdInsP2. These effects were not seen if isolated nuclei were incubated with IGF-I and bombesin. We interpret these results to mean that rapid mass changes occur in nuclear inositol phospholipids in the early stages of the mitotic response.

Animals↗

Rapid changes in phospholipid metabolism in the nuclei of Swiss 3T3 cells induced by treatment of the cells with insulin-like growth factor I.

When highly-purified nuclei of Swiss-mouse 3T3 cells are incubated with [32P]-gamma ATP, radioactivity is incorporated into phosphatidic acid and the two polyphosphoinositol lipids, phosphatidylinositol(4)P and phosphatidylinositol(4,5)P2. If the cells are pre-treated with insulin-like growth factor I, this incorporation into the phospholipids is decreased. The effect is maximal by 2 minutes, is transient in that it has disappeared by one hour, and is increased markedly by the co-addition of bombesin, even though bombesin alone has no effect. We suggest that some aspect of polyphosphoinositide metabolism is altered in the nucleus (leading to a decreased radiolabelling) when the cells are treated with mitogenic growth factors, and that this change in inositide metabolism is a very early event in the sequence leading to cell division.

Adenosine Triphosphate↗

Changes in ribonucleoprotein particle and chromatin organization induced by liposomes in isolated nuclei.

Nuclei isolated from rat liver, incubated in the presence of liposomes of different phospholipids, undergo typical modifications: chromatin dispersion and reduction of the interchromatin granules in nuclei incubated with negatively charged liposomes and increase of the chromatin density and of the number and size of the interchromatin granules in nuclei incubated with neutral liposomes. The possibility that the observed modifications are caused by an impairment of the transport and translocation of ribonucleoproteins belonging to the inner nuclear matrix, is suggested by the results obtained by radiotracer techniques on the release of RNA from liposome-incubated nuclei.

Animals↗

Evidence for a reduction of the replicative activity of matrix-bound DNA polymerase alpha following treatment with phospholipase C.

Since phospholipids have been suggested to play some role in the molecular organisation of the nuclear matrix, the effect of their removal by means of phospholipase C has been investigated in regenerating rat liver nuclear matrix. The matrix-bound DNA polymerase alpha shows an almost complete loss of activity following the digestion with phospholipase C. Since the polymerase activity is restored by adding exogenous DNA, we suggest that the effect is due to the removal of matrix residual DNA, which is in some way linked to the nuclear matrix by means of hydrophobic interactions.

Animals↗

Synthesis of polyphosphoinositides in nuclei of Friend cells. Evidence for polyphosphoinositide metabolism inside the nucleus which changes with cell differentiation.

Previous work demonstrated the existence of phosphatidylinositol kinase and phosphatidylinositol phosphate kinase in rat liver nuclei, with the suggestion that these activities are in the nuclear membrane [Smith & Wells (1983) J. Biol. Chem. 258, 9368-9373]. Here we show that highly purified nuclei from Friend cells, washed free of nuclear membrane by Triton, can incorporate radiolabel from [gamma-32P]ATP into phosphatidic acid, phosphatidylinositol phosphate and phosphatidylinositol 4,5-bisphosphate. The degree of radiolabelling of phosphatidylinositol bisphosphate is highly dependent on the state of differentiation of the cells, being barely detectable in growing cells and much greater after dimethyl sulphoxide-induced differentiation; this difference is mostly due to different amounts of phosphatidylinositol phosphate in the isolated nuclei. We suggest that polyphosphoinositides are made inside the nucleus and that they have a role in chromatin function; either the phospholipids themselves play a role, or there is a possibility of intranuclear signalling by inositide-derived molecules.

Animals↗

Ultrastructural organization of freeze-fractured interphase nuclei.

The morphology of intact or membrane-deprived interphase nuclei has been analysed by freeze-fracture electron microscopy. This method appears particularly useful for providing information on the distribution and organisation of chromatin and ribonucleoproteins in the absence of dehydration and embedding artifacts of conventional electron microscope techniques which, among other effects, appear to affect heterochromatin distribution, inducing its aggregation along the nuclear envelope. The main levels of chromatin superstructure, from nucleosome to solenoid fibres, are detectable in the replicas of freeze-fractured nuclei on the basis of the size of their shadow, a parameter particularly suitable for automated image analyses.

Animals↗

Uptake of tritiated phosphatidylcholine by isolated rat liver nuclei studied by electron microscope autoradiography in albumin embedded specimens.

The transfer of phosphatidylcholine from multilamellar vesicles to isolated rat liver nuclei was studied by means of electron microscope autoradiography. To avoid the possible loss or the artifactual redistribution of the phospholipid occurring during dehydration with organic solvents and plastic embedding, the fixed specimens were embedded in aqueous albumin, which was then hardened by glutaraldehyde and dehydrated physically. The quantitative analyses of the autoradiograms demonstrated that part of the labelled phosphatidylcholine was taken up by the isolated nuclei and was transferred inside the nucleoplasm. The uptake corresponded to the loss of the vesicular arrangement, probably owing to the formation of a lipoprotein complex with the nuclear proteins. The results provide evidence that the lipid-induced changes of transcriptional activity occur upon the actual interaction of the exogenous phospholipid with the inner nuclear components.

Albumins↗