TY - JOUR A1 - Franke, Werner W. A1 - Kleinschmidt, Jürgen A. A1 - Spring, Herbert A1 - Krohne, Georg A1 - Grund, Christine A1 - Trendelenburg, Michael F. A1 - Stöhr, Michael A1 - Scheer, Ulrich T1 - A nucleolar skeleton of protein filaments demonstrated in amplified nucleoli of Xenopus laevis N2 - The amplified, extrachromosomal nucleoli of Xenopus oocytes contain a meshwork of -4-nm-thick filaments, which are densely coiled into higher-order fibrils of diameter 30-40 nm and are resistant to treatment with high- and low-salt concentrations, nucleases (DNase I, pancreatic RNase, micrococcal nuclease), sulfhydryl agents, and various nonionic detergents. This filamentous "skeleton" has been prepared from manually isolated nuclear contents and nucleoli as weil as from nucleoli isolated by fluorescence-activated particle sorting. The nucleolar skeletons are observed in light and electron microscopy and are characterized by ravels of filaments that are especially densely packed in the nucleolar cortex. DNA as weil as RNA are not constituents of this structure, and precursors to ribosomal RNAs are completely removed from the extraction-resistant filaments by treatment with high-salt buffer or RN ase. Fractions of isolated nucleolar skeletons show specific enrichment of an acidic major protein of 145,000 mol wt and an apparent pi value of -6.15, accompanied in some preparations by various amounts of minor proteins. The demonstration of this skeletal structure in "free" extrachromosomal nucleoli excludes the problem of contaminations by nonnucleolar material such as perinucleolar heterochromatin normally encountered in studies of nucleoli from somatic cells. It is suggested that this insoluble protein filament complex forms a skeleton specific to the nucleolus proper that is different from other extraction-resistant components of the nucleus such as matrix and lamina and is involved in the spatial organization of the nucleolar chromatin and its transcriptional products. In studies of the organization of the interphase nucleus, considerable progress has been made in the elucidation of the arrangement of chromatin components and transcriptional products. However, relatively little is known about the composition and function of another category of nuclear structures, the nonnucleoproteinaceous architectural components that are insoluble in solutions of low and high ionic strength, despite numerous studies dedicated to this problem. Such structures include (a) the nuclear envelope and its pore complexes (I, 15, 18, 23, 37, 41), (b) a peripheral layer of insoluble protein ("lamina"; I, 15, 22, 23, 59), (e) certain skeletal proteins related to the chromosome "scaffold" described by Laemmli and coworkers (see references 2 and 3), and (d) ill-defined tangles of fibrillar structures of the nuclear interior that are collectively described as residual "matrix" (6, 21 ; for reviews, see references THE JOURNAL OF CEll BrOlOGY . VOlUME 90 AUGUST 1981 289-299 © The RockefeIler University Press · 0021 -9525/ 81 / 08/ 0289/ 11 $1 .00 4 and 12). The latter, preparatively Y1 - 1981 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-33130 ER - TY - JOUR A1 - Franke, Werner W. A1 - Scheer, Ulrich A1 - Trendelenburg, Michael F. A1 - Spring, Herbert A1 - Zentgraf, Hanswalter T1 - Absence of nucleosomes in transcriptionally active chromatin N2 - The ultrastructure of twO kinds of transcription ally active chromatin, the lampbrush chromosome loops and the nucleoli from amphibian oocytes and primary nuclei of the green alga Acetabularia, has been examined after manual isolation and dispersion in low salt media of slightly alkaline pH using various electron microscopic staining techniques (positive staining, metal shadowing, negative staining, preparation on positively charged films, etc.) and compared with the appearance of chromatin from various somatic cells (hen erythrocytes, rat hepatocytes, ClIltured murine sarcoma cells) prepared in parallel. While typical nucleosomes were revealed with all the techniques for chromatin from the latter three cell system, no nucleosomes were identified in either the lampbrush chromosome structures or the nucleolar chromatin. Nucleosomal arrays were absent not only in maximally fibril-covered matrix units but also in fibril-free regions between transcriptional complexes, including the apparent spacer intercepts between different transcriptional units. Moreover, comparisons of the length of the repeating units of rDNA in the transcribed state with those determined in the isolated rDNA and with the lengths of the first stable product of rDNA transcription, the pre-rRNA, demonstrated that the transcribed rDNA was not significantly shortened and/or condensed but rather extended in the transcriptional units. Distinct granules of about nucleosomal size which were sometimes found in apparent spacer regions as well as within matrix units of reduced fibril density were shown not to represent nucleosomes since their number per spacer unit was not inversely correlated with the length of the specific unit and also on the basis of their resistance to treatment with the detergent Sarkosyl NL-30. It is possible to structurally distinguish between transcriptionally active chromatin in which the DNA is extended in a non-nucleosomal form of chromatin and condensed, inactive chromatin within the typical nucleosomal package. The characteristic extended structure of transcriptionally active chromatin is found not only in the transcribed genes but also in non-transcribed regions within or between ("spacer") transcriptional units as well as in transcriptional units that are untranscribed amidst transcribed ones and/or have been inactivated for relatively short time. It is hypothesized that activation of transcription involves a transition from a nucleosomal to an extended chromatin organisation and that this structural transition is not specific for single "activated" genes but may involve larger chromatin regions, including adjacent untranscribed intercepts. KW - Cytologie KW - Chromatin structure KW - nucleosomes KW - transcription KW - electron microscopy Y1 - 1976 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-40646 ER - TY - JOUR A1 - Eckert, W. A. A1 - Franke, Werner W. A1 - Scheer, Ulrich T1 - Actinomycin D and the central granules in the nuclear pore complex: thin sectioning versus negative staining N2 - Thin section electron microscopy of Actinomycin D treated Tetrahymena cells and amphibian oocytes (Xenopus laevis, Triturus aZpestris) reveal no reduction in the central granules in the nuclear pore complexes. Possible reasons for the diversity between these results and earlier observations using negatively stained isolated nuclear envelopes from the same objects are discussed. The results clearly show that the presence of central granules within the nuclear pores does neither depend on nuclear RNA synthesis nor does indicate nucleocytoplasmic RNA transport. This conclusion leads to a reconsideration of the nature of the central granule. The functioning of the central granule of the nuclear pore complexes is further discussed in connection with recent studies on the ultrastructure of various types of cisternal pores. KW - Nuclear pores KW - Nucleocytoplasmic exchange KW - Actinomycin D KW - Tetrahymena KW - Amphibian oocytes Y1 - 1972 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-40636 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Franke, Werner W. T1 - Annulate lamellae in plant cells: formation during microsporogenesis and pollen development in Canna generalis Bailey N2 - The occurrence of stacked annulate tamellae is documented for a plant cell system, namely for pollen mother cells and developing pollen grains of Canna generalis. Their structural subarchiteeture and relationship to endoplasmie reticulum (ER) and nuclear envelope cisternae is described in detail. The results demonstrate structural homology between plant and animal annulate lamellae and are compatible with, though do not prove, the view that annulate lamcllar cisternae may originate as a degenerative form of endoplasmic retieulum. Y1 - 1972 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32160 ER - TY - JOUR A1 - Zentgraf, Hanswalter A1 - Scheer, Ulrich A1 - Franke, Werner W. T1 - Characterization and localization of the RNA synthesized in mature avian erythrocytes N2 - No abstract available Y1 - 1975 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32410 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Franke, Werner W. A1 - Trendelenburg, Michael F. A1 - Spring, Herbert T1 - Classification of loops of lampbrush chromosomes according to the arrangement of transcriptional complexes N2 - The arrangement of transcriptional units in the loops of lampbrush chromosomes from oocyte nuclei of urodele amphibia and from primary nuclei of the green alga Acetabularia have been studied in the electron microscope using spread preparations. Loops with different patterns of arrangement of matrix units (i.e. to a first approximation, transcriptional units) can be distinguished: (i) loops consisting of one active transcriptional unit; (ii) loops containing one active transcriptional unit plus additional fibril-free, i.e. apparently untranscribed, intercepts that may include 'spacer' regions; (iii) loops containing two or more transcriptional units arranged in identical or changing polarities, with or without interspersed apparent spacer regions. Morphological details of the transcriptional complexes are described. The observations are not compatible with the concept that one loop reflects one and only one transcriptional unit but, rather, lead to a classification of loop types according to the arrangement of their transcriptional units. We propose that the lampbrush chromosome loop can represent a unit for the coordinate transcription of either one gene or a set of several (different) genes. Y1 - 1976 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32822 ER - TY - JOUR A1 - Franke, Werner W. A1 - Jarasch, Ernst-Dieter A1 - Herth, Werner A1 - Scheer, Ulrich A1 - Zerban, Heide T1 - Cytology : general and molecular cytology N2 - The present review discusses some general aspects of membrane structure and problems of membrane isolation and membrane biochemistry, with particular focus on the endoplasmic reticulum. KW - Botanik Y1 - 1975 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41458 ER - TY - JOUR A1 - Franke, Werner W. A1 - Scheer, Ulrich A1 - Herth, Werner T1 - Cytology, general and molecular cytology N2 - The present article had originally been conceived as a review on endomembranes, the plasma membrane, and the major product of membrane-bound activities, the cell wall material. However, limitations of space and the cascading number of pertinent literature articles made it necessary to confine this to one group of membranes and one type of cell wall components. Therefore, we shall begin our survey on the biochemical and cytological aspects of membranes by a review of the class of the pore complex bearing endomembranes, i.e. the nuclear envelope and the annulate lamellae (AL). Next year the membranes of the endoplasmic reticulum and the dictyosomes will be dealt with in conjunction with a discussion of the various intracellular vesicles, the tonoplast and the plasmalemma. KW - Botanik Y1 - 1974 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-39499 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Franke, Werner W. A1 - Trendelenburg, Michael F. T1 - Effects of actinomycin D on the association of newly formed ribonucleoproteins with the cistrons of ribosomal RNA in Triturus oocytes N2 - No abstract available Y1 - 1975 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32383 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Kartenbeck, Jürgen A1 - Trendelenburg, Michael F. A1 - Stadler, Joachim A1 - Franke, Werner W. T1 - Experimental disintegration of the nuclear envelope: evidence for pore-connecting fibrils N2 - The disintegration of the nuclear envelope has been examined in nuclei and nuclear envelopes isolated from amphibian oocytes and rat liver tissue, using different electron microscope techniques (ultrathin sections and negatively or positively stained spread preparations). Various treatments were studied, including disruption by surface tension forces, very low salt concentrations, and non ionic detergents such as Triton X-lOO and Nonidet P-40. The high local stability of the cylinders of nonmembranous pore complex material is emphasized. As progressive disintegration occurred in the membrane regions, a network of fibrils became apparent which interconnects the pore complexes and is distinguished from the pore complexassociated intranuclear fibrils. This network might correspond to an indistinct lamella, about 15 - 20 nm thick, located at the level of the inner nuclear membrane, which is recognized in thin sections to bridge the interpore distances. With all disintegration treatments a somewhat higher susceptibility of the outer nuclear membrane is notable, but a selective removal does not take place. Final stages of disintegration are generally characterized by the absence of identifiable, membrane- like structures. Analysis of detergent-treated nuclei and nuclear membrane fractions shows almost complete absence of lipid components but retention of significant amount of glycoproteins with a typical endomembrane-type carbohydrate pattern. Various alternative interpretations of these observations are discussed. From the present observations and those of Aaronson and Blobel (1,2), we favor the notion that threadlike intrinsic membrane components are stabilized by their attachment to the pore complexes, and perhaps also to peripheral nuclear structures, and constitute a detergent-resistant, interpore skeleton meshwork. Y1 - 1976 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-39735 ER - TY - JOUR A1 - Trendelenburg, Michael F. A1 - Franke, Werner W. A1 - Scheer, Ulrich T1 - Frequencies of circular units of nucleolar DNA in oocytes of two insects, Acheta domesticus and Dytiscus marginalis, and changes of nucleolar morphology during oogenesis N2 - The organization of the extrachromosomal nucleolar material in oocytes of two insect species with different ovary types, the house cricket Acheta domesticus (panoistic ovary) and the water beetle Dytiscus marginalis (meroistic ovary), was studied with light and electron microscopic techniques. Stages early in oogenesis were compared with fully vitellogenic stages (mid-to-Iate diplotene). The arrangement of the nucleolar material undergoes a marked change from a densely aggregated to a dispersed state. The latter was characterized by high transcriptional activity. In spread and positively stained preparations of isolated nucleolar material, a high frequency of small circular units of transcribed rDNA was observed and rings with small numbers (1-5) of pre-rRNA genes were predominant. The observations suggest that the "extra DNA body" observed in early oogenic stages of both species represents a dense aggregate of numerous short circular units of nucleolar chromatin, with morphological subcomponents identifiable in ultrathin sections. These apparently remain in close association with the chromosomal nucleolar organizer(s). The observations further indicate that the individual small nucleolar subunit circles dissociate and are dispersed as actively transcribed rDNA units later in diplotene. The results are discussed in relation to principles of the ultrastructural organization of nucleoli in other cell types as well as in relation to possible mechanisms of gene amplification. KW - Zelldifferenzierung Y1 - 1977 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41370 ER - TY - JOUR A1 - Franke, Werner W. A1 - Spring, Herbert A1 - Scheer, Ulrich A1 - Zerban, Heide T1 - Growth of the nuclear envelope in the vegetative phase of the green alga Acetabularia. Evidence for assembly from membrane components synthesized in the cytoplasm. N2 - No abstract available Y1 - 1975 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32403 ER - TY - JOUR A1 - Trendelenburg, Michael F. A1 - Scheer, Ulrich A1 - Zentgraf, Hanswalter A1 - Franke, Werner W. T1 - Heterogeneity of spacer lengths in circles of amplified ribosomal DNA of two insect species, Dytiscus marginalis and Acheta domesticus N2 - No abstract available Y1 - 1976 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-33055 ER - TY - JOUR A1 - Kleinschmidt, Jürgen A. A1 - Scheer, Ulrich A1 - Dabauvalle, Marie-Christine A1 - Bustin, Michael A1 - Franke, Werner W. T1 - High mobility group proteins of amphibian oocytes: a large storage pool of a soluble high mobility group-1-like protein and involvement in transcriptional events N2 - No abstract available Y1 - 1983 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-33250 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Messner, Karin A1 - Hazan, Rachel A1 - Raska, Ivan A1 - Hansmann, Paul A1 - Falk, Heinz A1 - Spiess, Eberhard A1 - Franke, Werner W. T1 - High sensitivity immunolocalization of double and single-stranded DNA by a monoclonal antibody N2 - A monoclonal antibody (AK 30-10) is described which specifically reacts with DNA both in double and single-stranded forms but not with other molecules and structures, including deoxyribonucleotides and RNAs. When used in immunocytochemical experiments on tissue sections and permeabilized cultured cells, this antibody detects DNA-containing structures, even when the DNA is present in very small amounts. Examples of high resolution detection include the DNA present in amplified extrachromosomal nucleoli, chromomeres of lampbrush chromosomes, mitochondria, chloroplasts and mycoplasmal particles. In immunoelectron microscopy using the immunogold technique, the DNA was localized in distinct substructures such as the "fibrillar centers" of nucleoli and certain stromal centers in chloroplasts. The antibody also reacts with DNA of chromatin of living cells, as shown by microinjection into cultured mitotic cells and into nuclei of amphibian oocytes. The potential value and the limitations of immunocytochemical DNA detection are discussed. KW - Cytologie KW - DNA antibodies KW - monoclonal antibodies KW - DNA immunolocalization KW - chromatin KW - mycoplasma tests Y1 - 1987 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41063 ER - TY - JOUR A1 - Spring, Herbert A1 - Krohne, Georg A1 - Franke, Werner W. A1 - Scheer, Ulrich A1 - Trendelenburg, Michael F. T1 - Homogeneity and heterogeneity of sizes of transcriptional units and spacer regions in nucleolar genes of Acetabularia N2 - The arrangement of genes of precursor molecules for ribosomal RNA (pre-rRNA) in primary nuclei from two green algae species, Acetabularia mediterranea and A. major, has been analyzed in an electron microscope study. The pattern of transcriptional units in individual strands of nucleolar chromatin was investigated using spread and positively stained preparations. The rDNA pattern is not uniform but differs in different strands. The predominant type of nucleolar chromatin exhibits a high degree of homogeneity in the sequence of matrix units (intercepts covered with fibrilst hat contain the pre-rRNA) and fibril-free spacer intercepts. Substantial differences, however, are observed between the patterns in different strands. In addition, there is evidence in some strands for intraaxial heterogeneity of both spacer and matrix units. The following major types can be distinguished: type la, ca. 2 micrometer long matrix units, extremely short spacer intercepts in A. mediterranea (ca. 1 micrometer long ones in A. major), completely homogeneous distribution; type Ib, as type la but with intercalated, isolated, significantly shorter and/or longer matrix units; type lIa, matrix unit sizes as in type la, but much longer spacer intercepts, high degree of homogeneity; type Ill, largely heterogeneous arrangements of matrix and spacer units of varying sizes. The matrix unit data are compared with the sizes of pre-rRNA as determined by polyacrylamide gelelectrophoresis under denaturing and non-denaturing conditions. The findings are discussed in relation to recent observations in amphibia and insects and with respect to current concepts of the species-specificity of rDNA arrangements. Y1 - 1976 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41398 ER - TY - JOUR A1 - Scheer, Ulrich A1 - Schmidt-Zachmann, Marion S. A1 - Hügle, Barbara A1 - Franke, Werner W. T1 - Identification and localization of a novel nucleolar protein of a high molecular weight by a monoclonal antibody N2 - A monoclonal murine antibody (No-I 14) is described which reacts specifically with a polypeptide of molecular weight (M,) 180000 present in low-speed nuclear pellets from oocytes and somatic cells of Xenopus laevis and X. borealis and in isolated amplified nucleoli. Two-dimensional gel electrophoresis has revealed the acidic nature of this polypeptide (isoelectric at pH of ca 4.2 in the presence of 9.5 M urea). A relatively large proportion of the protein is extracted at elevated ionic strength( i.e., at 0.4-0.5 M alkali salt) in a form sedimenting at approx. 7-8S , compatible with a monomeric state. It is also extracted by digestion with RNase but not with DNase. In immunofluorescence microscopy, antibody No-114 stains intensely nucleoli of oocytes and all somatic cells examined , including the residual nucleolar structure of Xenopus erythrocytes which are transcriptionally inactive. During mitosis the antigen does not remain associated with the nucleolar organizer regions (NOR) of chromosomes but is released and dispersed over the cytoplasm until telophase when it re-associates with the reforming interphase nucleoli. At higher resolution the immunofluorescent region is often resolved into a number of distinct subnucleolar components of varied size and shape. Immunoelectron microscopy using colloidal gold-coupled secondary antibodies reveals that the M, 180000 protein is confined to the dense fibrillar component of the nucleolus. This conclusion is also supported by its localization in the fibrillar part of segregated nucleoli of cells treated with actinomycin D. We conclude that nucleoli contain a prominent protein of M, 180000 which contributes to the general structure of the dense fibrillar component of the interphase nucleolus , independent of its specific transcriptional activity. Y1 - 1984 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-39786 ER - TY - JOUR A1 - Weber, Klaus A1 - Osborn, Mary A1 - Franke, Werner W. A1 - Seib, Erinita A1 - Scheer, Ulrich A1 - Herth, Werner T1 - Identification of microtubular structures in diverse plant and animal cells by immunological cross-reaction revealed in immunofluorescence microscopy using antibodies against tubulin from porcine brain N2 - Antibody against tubulin from porcine brain was used to evaluate the immunological cross reactivity of tubulin from a variety of animal and plant cells. Indirect immunofluorescence microscopy revealed microtubule-containing structures including cytoplasmic microtubules, spindle microtubules, cilia and fIagella. Thus tubulin from diverse species of both mammals and plants show immunological cross-reactivity with tubulin from porcine brain. Results obtained by immunofluorescence microscopy are whenever possible compared with previously known ultrastructural results obtained by electron microscopy. KW - Cytologie KW - Microtubules KW - immunofluorescence KW - evolution KW - antibody KW - sperm Y1 - 1977 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-41383 ER - TY - JOUR A1 - Franke, Werner W. A1 - Kartenbeck, Jürgen A1 - Krien, S. A1 - VanderWoude, W. J. A1 - Scheer, Ulrich A1 - Morré, D. J. T1 - Inter- and intracisternal elements of the Golgi apparatus: A system of membrane-to-membrane cross-links N2 - Electron opaque cross-bridge structures span the inter- and intracisternal spaces and provide membrane-to-membrane connections between adjacent cisternae of dictyosomes of pollen tubes of Clivia and Lilium. Additionally, the classic intercisternal rods, characteristic of intercisternal regions near the maturing face of dictyosomes, are connected with the adjacent membranes through similar cross-bridge elements. We suggest that these structural links are responsible for maintaining the flattened appearance of the central parts of Golgi apparatus cisternae as well as for the coherence of cisternae within the stack. Observations on other plant (e.g. microsporocytes of Canna) and animal cells (e.g. rodent liver and hepatoma cells, newt spermatocytes) show that such an array of membrane cross-links is a universal feature of Golgi apparatus architecture. The cross-bridges appear as part of the complex "zone of exclusion" which surrounds dictyosomes, entire Golgi apparatus and Golgi apparatus equivalents in a variety of cell types. KW - Golgi apparatus KW - Membranes KW - Cross-bridges KW - Electron microscopy Y1 - 1972 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-39514 ER - TY - JOUR A1 - Franke, Werner W. A1 - Scheer, Ulrich A1 - Fritsch, Hansjörg T1 - Intranuclear and cytoplasmic annulate lamellae in plant cells N2 - No abstract available Y1 - 1972 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-32148 ER -