@article{ThiryScheerGoessens1988, author = {Thiry, Marc and Scheer, Ulrich and Goessens, Guy}, title = {Immunoelectron microscopic study of nucleolar DNA during mitosis in Ehrlich tumour cells}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40745}, year = {1988}, abstract = {In order to investigate the DNA localization within Ehrlich tumor cell nucleoli during mitosis, two recent immunocytochemical methods using either an anti-DNA or an anti-bromodeoxyuridine (BrdU) monoclonal antibody have been applied. In both cases, the immunogold labeling has been performed on ultrathin sections of cells embedded either in Lowicryl K4M or in Epon, respectively. Identical results are observed with both immunocytochemical approaches. In the interphase nucleolus, besides the labeling of the perinucleolar chromatin shell and of its intranucleolar invaginations which penetrate into the nucleolar body and often terminate at the fibrillar centers, a few gold particles are also preferentially found towards the peripheral region of the fibrillar centers. In contrast, the dense fibrillar component and the granular component are never labeled. During mitosis, the fibrillar centers persist at the chromosomal nucleolus organizing regions (NOR's) and can be selectively stained by the silver method. However, these metaphase fibrillar centers are no longer decorated by the DNA- or BrdU antibodies. These results indicate that until the end of prophase, rRNA genes are present inside the fibrillar center material, disappear during metaphase and reappear in reconstituting nucleoli during telophase. Thus, fibrillar centers appear to represent structures sui generis, which are populated by rRNA genes only when the nucleolus is functionally active. In segregated nucleoli after actinomycin D treatment, the DNA labeling is exclusively restricted to the perinucleolar chromatin blocks. These findings also suggest that the DNA content of the fibrillar center material varies according to the rRNA transcription level of the cells. The results are discussed in the light of the present knowledge of the functional organization of the nucleolus.}, subject = {Cytologie}, language = {en} } @article{RoseSzopaHanetal.1988, author = {Rose, Kathleen M. and Szopa, Jan and Han, Fu-Sheng and Cheng, Yung-Chi and Richter, Arndt and Scheer, Ulrich}, title = {Association of DNA topoisomerase I and RNA polymerase I: A possible role for topoisomerase I in ribosomal gene transcription}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-33901}, year = {1988}, abstract = {RNA polymerase I preparations purified from a rat hepatoma contained DNA topoisomerase activity. The DNA topoisomerase associated with the polymerase had an Mr of 110000, required Mg2+ but not ATP, and was recognized by anti-topoisomerase I antibodies. When added to RNA polymerase I preparations containing topoisomerase activity, anti-topoisomerase I antibodies were able to inhibit the DNA relaxing activity of the preparation as well as RNA synthesis in vitro. RNA polymerase II prepared by analogous procedures did not contain topoisomerase activity and was not recognized by the antibodies. The topoisomerase I: polymerase I complex was reversibly dissociated by column chromatography on Sephacryl S200 in the presence of 0.25 M (NH4hS04. Topoisomerase I was immunolocalized in the transcriptionally active ribosomal gene complex containing RNA polymerase I in situ. These data indicate that topoisomerase I and RNA polymerase I are tightly complexed both in vivo and in vitro, and suggest a role for DNA topoisomerase I in the transcription of ribosomal genes.}, language = {en} } @article{ReimerRaskaScheeretal.1988, author = {Reimer, Georg and Raska, Ivan and Scheer, Ulrich and Tan, Eng M.}, title = {Immunolocalization of 7-2-ribonucleoprotein in the granular component of the nucleolus}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-33890}, year = {1988}, abstract = {Certain autoimmune sera contain antibodies against a nucleolar ribonucleoprotein particle associated with 7-2-RNA (R. Reddy et al. (1983) J. Bioi. Chem . 258, 1383; C. Hashimoto and J. A. Steitz (1983) J. Bioi. Chem. 258, 1379). In this study, we showed by immunofluorescence microscopy that antibodies reactive with 7-2-ribonucleoprotein immunolocalized in the granular regions of actinomycin D and 5,6-dichloro-I-j3-D-ribofuranosylbenzimidazole (DRB)-segregated nucleoli from Vero cells. By electron microscopic immunocytochemistry, antigen-antibody complexes were located in the granular component of transcriptionally active nucleoli from rat liver hepatocytes and HeLa cells. Anti-7- 2-RNP antibodies from two autoimmune sera immunoprecipitated a major protein of Mr 40,000 from e5S] methionine-Iabeled HeLa cell extract. The immunolocalization data suggest that 7-2-ribonucleoprotein may be involved in stages of ribosome biogenesis which take place in the granular component of the nucleolus, i.e., assembly, maturation, and/or transport of preribosomes}, language = {en} } @article{BenaventeSchmidtZachmannHuegleDoerretal.1988, author = {Benavente, Ricardo and Schmidt-Zachmann, Marion S. and H{\"u}gle-D{\"o}rr, B. and Reimer, G. and Rose, K. M. and Scheer, Ulrich}, title = {Identification and definition of nucleolus-related fibrillar bodies in micronucleated cells}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-39423}, year = {1988}, abstract = {Small nucleolus-related bodies which occur in the nUcleoplasm of " micronuclei" lacking nucleolar organizers have been studied by immunofluorescence microscopy. These bodies stained specifically with three different antibodies directed against proteins that are normally associated with the dense fibrillar component of functional nucleoli, but not with antibodies specific for certain proteins of the granular component or the fibrillar centers. Our data show that, in the absence of rRNA genes, the various constituent proteins characteristic of the dense fibrillar component spontaneously assemble into spherical entities but that the subsequent fusion of these bodies into larger structures is prevented in these micronuclei. The similarity between these nucleolus-related bodies of micronuclei and the prenucleolar bodies characteristic of early stages of nucleologenesis during mitotic telophase is discussed.}, language = {en} } @article{ThiryScheerGoessens1988, author = {Thiry, Marc and Scheer, Ulrich and Goessens, Guy}, title = {Localization of DNA within Ehrlich tumour cells nucleoli by immunoelectron microscopy}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-39327}, year = {1988}, abstract = {The distribution of DNA in Ehrlich tumour cell nucleoli was investigated by means of an immunocytochemical approach , involving a monoclonal antibody directed against double- and single-stranded DNA. Immunolabelling was performed . either before or after the embedding process. The postembedding labelling method allows better ultrastructural preservation than the preembedding labelling method. In particular, the various nucleolar components are well preserved and identifiable. In the nucleolus, labelling is particularly concentrated over the perinucleolar chromatin and over its intranucleolar invaginations, which penetrate the nucleolar body and often terminate at the fibrillar centres. In addition, aggregates of gold particles are found in the fibrillar centres, preferentially towards the peripheral regions. By contrast, the dense fibrillar component is completely devoid of labelling. The results seem to indicate that DNA containing the rDNA genes is located in the fibrillar centres, with a preference for the peripheral regions. This finding suggests that transcription of the rDNA genes should occur within the confines of the fibrillar centre, probably close to the boundary region of the surrounding dense fibrillar component. The results are discussed in the light of present knowledge of the functional organization of the nucleolus.}, language = {en} } @article{DabauvalleSchulzScheeretal.1988, author = {Dabauvalle, Marie-Christine and Schulz, Barbara and Scheer, Ulrich and Peters, Reiner}, title = {Inhibition of nuclear accumulation of karyophilic proteins in living cells by microinjection of the lectin wheat germ agglutinin}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-34288}, year = {1988}, abstract = {No abstract available}, language = {en} } @article{ScheerDabauvalleMerkertetal.1988, author = {Scheer, Ulrich and Dabauvalle, Marie-Christine and Merkert, Hilde and Benavente, Ricardo}, title = {The nuclear envelope and the organization of the pore complexes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-34275}, year = {1988}, abstract = {No abstract available}, language = {en} } @article{BenaventeReimerRoseetal.1988, author = {Benavente, Ricardo and Reimer, Georg and Rose, Kathleen M. and H{\"u}gle-D{\"o}rr, Barbara and Scheer, Ulrich}, title = {Nucleolar changes after microinjection of antibodies to RNA polymerase I into the nucleus of mammalian cells}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40666}, year = {1988}, abstract = {After microinjection of antibodies against RNA polymerase I into the nuclei of cultured rat kangaroo (PtKz) and rat (RVF-SMC) cells alterations in nucleolar structure and composition were observed. These were detected by electron microscopy and double-label immunofluorescence microscopy using antibodies to proteins representative of the three major components of the nucleolus. The microinjected antibodies produced a progressive loss of the material of the dense fibrillar component (DFC) from the nucleoli which, at 4 h after injection, were transformed into bodies with purely granular component (GC) structure with attached fibrillar centers (FCs). Concomitantly, numerous extranucleolar aggregates appeared in the nucleoplasm which morphologically resembled fragments of the DFC and contained a protein (fibrillarin) diagnostic for this nucleolar structure. These observations indicate that the topological distribution of the material constituting the DFC can be experimentally influenced in interphase cells, apparently by modulating the transcriptional activity of the rRNA genes. These effects are different from nucleolar lesions induced by inhibitory drugs such as actinomycin D-dependent "nucleolar segregation". The structural alterations induced by antibodies to RNA polymerase I resemble, however, the initial events of nucleolar disintegration during mitotic prophase.}, language = {en} }