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D Paulin

Publications and source records attributed to D Paulin.

At least 127 records · Page 7Linked to original sources

Cytoskeleton organization in differentiating mouse teratocarcinoma cells.

In addition to the well-known cytoskeleton actin and tubulin, new intermediate-sized filaments have been isolated that ensure cellular connections. Their proteins, referred to here as IFP (intermediate filament proteins), comprise keratin desmin, vimentin, glialin and neurofilin. We review some of these new findings, with emphasis on the IFP expressions in the differentiation of embryonic cells and the deviations encountered in tumor cells. The first measurements on embryonic and differentiated mouse teratoma carcinoma cells established that all cell lines contain cytoplasmic microtubules. Although actin is also present in all cells, its organization differs markedly in differentiated derivatives. Since then, extensive studies have confirmed these findings and refined our understanding of the adaptation of cytoskeletons during differentiation to fulfill thier function. Fibrillar IFP structures have diameters ranging from 80 to 150 A, by electromicroscopic measurements. The specificity established in various structural functions have emerged from protein assays by two-dimensional electrophoresis and by immunofluorescence. With mouse teratoma cells, the incipient formation of the three structural components is followed by applying antibodies of high specificity for keratin, desmin and vimentin. During in vitro differentiation of teratoma carcinoma cells, as in animal tissue, keratin is expressed always in the endodermal cells and desmin in the muscle cells. Vimentin, which is known to be restricted to mesenchymal tissue in animals, occurs in all cells which have acquired the potential for unlimited growth in culture. In embryonic developments, all cells of mouse blastocyst express microtubules which is consistent with the function attributed to tubulin. The structure of actin, by contrast, changes during the first morphological step of differentiation. The outer trophectodermal cells contain actin cables, whereas the inner cell mass contains actin in a diffuse state. It has been established that keratin fibers appear in trophoblastic cells, i.e., in the earliest embryonic differentiation of epithelial character. Research on the development of IFP is rapidly progressing. Recent results of several groups are discussed.

Actins↗

Reorganization of the cytoskeleton by interferon in MSV-transformed cells.

beta and gamma interferons were found to enhance the organization of microfilaments, intermediate filaments, and fibronectin in the murine sarcoma virus (MSV)-transformed cell line. Furthermore, both interferons increased the number of intermediate-type junctions. Sodium butyrate was also found to act in a similar way and, when associated with interferon, an additive effect was observed. In the cell line, cultured for many passages in the presence of beta interferon (MSV-IF+), the cytoskeletal network and the extra-cellular matrix were highly developed and were reminiscent of a normal cell. These results could explain some of the biological actions of interferon, such as increased cell adhesion to the substratum and decreased cell motility.

Actins↗

A comparison of surface proteins in embryonal carcinoma cells and their differentiated derivatives.

Surface proteins from five cell lines (three embryonal carcinoma cell lines (F9, PCC4 and PCC3), teratocarcinoma-derived endodermal cells (PYS) and fibroblasts (line 3/A/1-D-3 differentiated from PCC3) were compared by two-dimensional polyacrylamide gel electrophoresis after selective iodination with 125I in the presence of lactoperoxidase. The labeled proteins were solubilized either in Nonidet P40/urea/ampholyte/mercaptoethanol solution or in Nonidet P40 only. In total, about thirty major 125I-labeled surface proteins were identified by their isoelectric point and molecular weight. 14 proteins are present in all five cell types, although their quantity or accessibility for labeling differs between differentiated and undifferentiated cells. Three proteins (200, 160 and 150 kilodaltons) are present in undifferentiated cells only. Two of them (160 and 150 kilodaltons) were solubilized by Nonidet P40/urea/ampholyte/mercaptoethanol, but not by Nonidet P40. One protein (50 kilodaltons) was found in nullipotent F9 cells only. About 14--15 proteins (including fibronectin) were released by Nonidet P40/urea/ampholyte/mercaptoethanol but not by Nonidet P40. They are presumably bound to submembrane or cytoskeleton structures by non-covalent bonds.

Animals↗

Tropomyosin synthesis accompanies formation of actin filaments in embryonal carcinoma cells induced to differentiate by hexamethylene bisacetamide.

Hexamethylene bisacetamide (HMBA) induces in vitro the cytodifferentiation of PCC3/A/1 mouse embryonal carcinoma (EC) cells. In EC cells, actin is associated with surface structures but microfilament bundles are not seen. After 2 days of HMBA treatment, rounded EC cells are converted to flat adhesive ones with a developed cytoskeleton containing actin and tropomyosin. The ratio of actin to total proteins is constant in EC cells and their HMBA derivatives; but a striking difference is observed for one of the newly synthesized proteins (Mr 34,000) identified as tropomyosin. Synthesis of tropomyosin is followed by its association with actin microfilament bundles, as revealed by indirect immunofluorescence microscopy with specific antibodies.

Acetamides↗

[Expression of ts-3 mutant of polyoma virus in mouse cells (author's transl)].

A thermosensitive mutant (ts-3) of polyoma virus produces T antigen, viral DNA and V antigen in BalbC/3T3 cells only at the permissive temperature (31 degrees C). At the non-permissive temperature (39 degrees C) these cells are unable to complement the viral defect. The pulse technique reveals only transitory cellular DNA induction between 15 and 25 h after infection at 39 degrees C. However, at both temperatures, cytoplasmic penetration is observed, and the viral DNA reaches the nuclei. Similarly, the myoblast cell line PCD2 does not complement the viral defect. In other idfferent types of cells tested (3T6, mouse kidney or secondary embryo cultures), the ts-3 mutant is expressed equally at 31 degrees C and 39 degrees C.

Animals↗

Immunofluorescence studies on deoxyribonuclease from mouse teratocarcinoma cells during cell differentiation.

Specific anti-DNase-I IgG have been used to detect deoxyribonuclease in teratocarcinoma cells by an indirect immunofluorescence method. All the cells studied show fluorescence staining. However, the patterns are quite different in embryonal carcinoma cells (amorphous cytoplasmic fluorescence and absence of nuclear staining) as compared to differentiated cell lines (diffuse, bright granular nuclear and fibrillar cytoplasmic fluorescence). It is possible by this method to distinguish different cell types derived from the same origin. Deoxyribonuclease from teratocarcinoma cells can therefore be considered as a marker of cell differentiation in this system.

Animals↗

Polyoma virus T antigen. I. Synthesis of modified heat-labile T angiten in cells transformed with the ts-a mutant.

In mouse cells transformed with the ts-a mutant of polyoma virus (ts-a-3T3), only low amounts of the virus-specific T antigen were synthesized at high temperature (39 C). After a shift-down to the permissive temperature (31 C), these cells exhibited the same level of T-antigen production as wild-type polyoma transformants. The T antigen produced by ts-a-transformed cells was inactivated at 39 C in vitro at a faster rate than that produced by wild-type-transformed cells. These observations indicate that T antigen is, or includes, a virus-coded peptide.

Animals↗

Immunochemical procedure for partial purification of newly synthesized proteins in polyoma virus-infected mouse cells.

A broad-spectrum antiserum has been obtained against nuclear proteins from normal mouse fibroblasts. Solid phase immunoadsorbants prepared from the serum specifically bind the corresponding antigens. The removal in this way of a significant fraction of host proteins leads to partial purification of viral and virus-induced polypeptides from polyoma virus-infected cell extracts. Four main peptides can be selected, with respective molecular weights of 90,000, 70,000, 46,000, and 41,000.

Adsorption↗