Biomedical subjects

G Tabellini

Publications and source records attributed to G Tabellini.

23 records · Page 2Linked to original sources

PubMed1999

Influence of different metal ions on the ultrastructure, biochemical properties, and protein localization of the K562 cell nuclear matrix.

The higher order of chromatin organization is thought to be determined by the nuclear matrix, a mainly proteinaceous structure that would act as a nucleoskeleton. The matrix is obtained from isolated nuclei by a series of extraction steps involving the use of high salt and nonspecific nucleases, which remove chromatin and other loosely bound components. It is currently under debate whether these structures, isolated in vitro by unphysiological extraction buffers, correspond to a nucleoskeleton existing in vivo. In most cell types investigated, the nuclear matrix does not spontaneously resist these extractions steps; rather, it must be stabilized before the application of extracting agents. In this study nuclei, isolated from K562 human erythroleukemia cells, were stabilized by incubation with different metal ions (Ca2+, Cu2+, Zn2+, Cd2+), and the matrix was obtained by extraction with 2 M NaCl. By means of ultrastructural analysis of the resulting structures, we determined that, except for Ca2+, all the other metals induced a stabilization of the matrix, which retained the inner fibrogranular network and residual nucleoli. The biochemical composition, analyzed by two-dimensional gel electrophoresis separation, exhibited a distinct matrix polypeptide pattern, characteristic of each type of stabilizing ion employed. We also investigated to what extent metal ions could maintain in the final structures the original distribution of three inner matrix components, i.e. NuMA, topoisomerase IIalpha, and RNP. Confocal microscopy analysis showed that only NuMa, and, to a lesser extent, topoisomerase IIalpha, were unaffected by stabilization with divalent ions. On the contrary, the fluorescent RNP patterns detected in the resulting matrices were always disarranged, irrespective of the stabilization procedure. These results indicate that several metal ions are powerful stabilizing agents of the nuclear matrix prepared from K562 erythroleukemia cells and also strengthen the concept that NuMA and topoisomerase IIalpha may act as structural components of the nuclear matrix.

Antigens, Neoplasm↗

PubMed1999

Insulin-like growth factor-I-dependent stimulation of nuclear phospholipase C-beta1 activity in Swiss 3T3 cells requires an intact cytoskeleton and is paralleled by increased phosphorylation of the phospholipase.

Swiss 3T3 mouse fibroblasts were exposed to 10 microM colchicine to disrupt microtubules, then stimulated with insulin-like growth factor-I. Immunoprecipitation experiments showed that insulin-like growth factor-I receptor and insulin receptor substrate-1 were tyrosine phosphorylated to the same extent in both cells treated with colchicine and in those not exposed to the drug. Moreover, the activity of phosphatidylinositol 3-kinase was not affected by incubation with colchicine. While in nuclei prepared from cells not exposed to colchicine it was possible to detect an insulin-like growth factor-I-dependent increase in the mass of diacylglycerol, as well as stimulation of phospholipase C activity, no similar changes were observed in nuclei obtained from cells treated with colchicine. Activation of the nuclear phospholipase activity was paralleled by an increase of its phosphorylation. Immunofluorescent studies revealed that mitogen-activated protein kinase did not translocate towards the nucleus when the cytoskeleton was depolymerized. These results show that in Swiss 3T3 cells some as yet unknown events necessary for the insulin-like growth factor-I-dependent activation of nuclear polyphosphoinositide metabolism require the presence of an intact cytoskeleton and are situated down-stream the activation of insulin receptor substrate-1 and phosphatidylinositol 3-kinase. Activation of nuclear phospholipase C-beta1 might be linked to its phosphorylation and translocation of mitogen-activated protein kinase to the nucleus.

3T3 Cells↗

PubMed1998

Intranuclear distribution of HMGI/Y proteins. An immunocytochemical study.

The intranuclear distribution of HMGI/Y proteins was analyzed by immunofluorescent staining in several cell lines using a polyclonal antibody that stained a fibrogranular network. In actively growing 3T3 fibroblasts, HMGI/Y proteins were mainly localized to heterochromatin masses, whereas in quiescent cells they were more diffusely distributed. Double labeling experiments showed a co-localization of HMGI/Y with DNA topoisomerase IIalpha. These results are in agreement with previously published biochemical data and indicate a possible involvement of HMGI/Y proteins in several nuclear functions, including chromatin organization and gene expression.

3T3 Cells↗

PubMedDate not supplied

Further considerations on the intranuclear distribution of HMGI/Y proteins.

We have investigated the intranuclear distribution of High-mobility group proteins I/Y by means of immunofluorescent staining employing a variety of cell lines. Confocal scanning laser microscopy analysis revealed that High-mobility group proteins I/Y are present in an intranuclear fibrogranular network and mitotic chromosomes. In Caski, C4I, Flow 2002, and K562 cell lines, High-mobility group proteins I/Y were absent from nucleoli, whereas in HeLa cells they were present in this nuclear domain. Double immunolabeling studies showed that High-mobility group proteins I/Y co-localize with other key nuclear components such as NuMA, SC-35, and TAF(II)70. Nuclear reactivity for High-mobility group proteins I/Y dramatically decreased in apoptotic nuclei of HL-60 human leukemia cells. Our morphological data correlate well with previous biochemical and functional findings obtained by other investigators, who have demonstrated multiple roles for this class of polypeptides. However, they point to the likelihood that High-mobility group proteins I/Y are involved in as yet unidentified functions, most likely in the speckle domains of the nucleus. They also show that conceivably these proteins are also involved in apoptosis.

Antigens, Nuclear↗

PubMedDate not supplied

Nuclear expression of diacylglycerol kinases: possible involvement in DNA replication.

The existence of intranuclear lipid-dependent signal transduction systems has been demonstrated by several independent groups. Remarkably, intranuclear lipid-dependent signal transduction pathways are regulated independently from their membrane/cytosolic counterparts. A sizable body of evidence suggests that nuclear lipid signaling controls critical biological functions such as cell proliferation, differentiation, and apoptosis. Diacylglycerol (DG) is a fundamental lipid second messenger which is produced in the nucleus. Since the levels of nuclear DG fluctuate during the cell cycle progression, it has been suggested that this lipid second messenger has important regulatory roles. Most likely, nuclear DG serves as a chemoattractant for some isoforms of protein kinase C that migrate to the nucleus in response to a variety of agonists. The nucleus also contains diacylglycerol kinases (DGKs), i.e. the enzymes that, by converting DG into phosphatidic acid (PA), terminate DG-dependent events. This review aims at highlighting the different isozymes of DGKs present within the nucleus as well as at discussing their potential functions with particular emphasis placed on DNA replication.

Animals↗