dThd bound to epoxy-activated Sepharose results in a high specific affinity adsorbent, which was ... more dThd bound to epoxy-activated Sepharose results in a high specific affinity adsorbent, which was used to purify Physarum dThd kinase 8000-fold. With this purified enzyme, it was proved that dThd kinase exists in five enzyme variants in Physarum polycephalum, the acidic forms being phosphorylated. This protein modification is discussed under the aspect of the regulation of this enzyme.
Lamin-like proteins have been identified as components of the nuclear matrix of the myxomycete Ph... more Lamin-like proteins have been identified as components of the nuclear matrix of the myxomycete Physarum polycephalum. A 67 kDa homologue was detected by immunoblotting of nuclear matrix proteins with affinity-purified anti-lamin antibodies of human autoimmune sera. A 65 kDa lamin B homologue was identified with polyclonal antibodies against turkey erythrocyte lamin B in nuclei and nuclear matrix of Physarum amoebae and plasmodia. Indirect immunofluorescence microscopy revealed that the 65 and 67 kDa proteins are localized in the nuclear lamina of plasmodial nuclei.
Interferon regulatory factor (IRF)-1 expression was surveyed in nontransformed and oncogene-trans... more Interferon regulatory factor (IRF)-1 expression was surveyed in nontransformed and oncogene-transformed mouse fibroblasts, using Western immunoblot with an IRF-1-specific antiserum, to examine possible differences resulting from cellular transformation. Ten additional proteins that reacted with the IRF-1 antibody and that underwent specific competition by peptide antigen were observed in extracts of both nontransformed and oncogene-transformed cell lines. Cross-reacting proteins were also observed in mouse macrophage extracts. Protein was captured from fibroblast nuclear extracts, using oligonucleotides representing IRF-binding sequences linked to magnetic beads. Captured proteins were eluted and analyzed by immunoblot with anti-IRF-1. Along with 43-kDa IRF-1, 4 of the 7 nuclearly located cross-reacting proteins (97, 90, 66, and 33 kDa) were found to complex with the IRF binding element. These proteins, with an epitope in common with the IRF-1 C-terminal region and IRF element DNA sequence-binding capability, may represent new members of the IRF family.
... J. 303, 723−729. Sendra, R., Rodrigo, I., Salvador, ML, & Franco, L. (1988) Plant... more ... J. 303, 723−729. Sendra, R., Rodrigo, I., Salvador, ML, & Franco, L. (1988) Plant Mol. Biol. 11, 857−866. ... Antonello Mai, Silvio Massa, Dante Rotili, Silvia Simeoni, Rino Ragno, Giorgia Botta, Angela Nebbioso, Marco Miceli, Lucia Altucci, and Gerald Brosch. ...
Hintergrund: Halsschmerzen sind eine der häufigsten Konsultationsursachen in der AM-Praxis. Bei K... more Hintergrund: Halsschmerzen sind eine der häufigsten Konsultationsursachen in der AM-Praxis. Bei Kindern sind ca. 20% und bei Erwachsenen <5% ß-hämolysierende Streptokokken der Gruppe A zu erwarten. 75% der Erwachsenen verlassen die Ordination aber mit einer Antibiotikaverordnung.[for full text, please go to the a.m. URL]
The readout of the genetic information of eukaryotic organisms is significantly regulated by modi... more The readout of the genetic information of eukaryotic organisms is significantly regulated by modifications of DNA and chromatin proteins. Chromatin alterations induce genome-wide and local changes in gene expression and affect a variety of processes in response to internal and external signals during growth, differentiation, development, in metabolic processes, diseases, and abiotic and biotic stresses. This review aims at summarizing the roles of histone H1 and the acetylation and methylation of histones in filamentous fungi and links this knowledge to the huge body of data from other systems. Filamentous fungi show a wide range of morphologies and have developed a complex network of genes that enables them to use a great variety of substrates. This fact, together with the possibility of simple and quick genetic manipulation, highlights these organisms as model systems for the investigation of gene regulation. However, little is still known about regulation at the chromatin level i...
Postsynthetic acetylation of core histones exhibits a peak during S-phase of the Physarum cell cy... more Postsynthetic acetylation of core histones exhibits a peak during S-phase of the Physarum cell cycle. The maximum H-acetate Incorporation precedes the maximum of hi stone synthesis. Acetate is incorporated into all core histones during S-phase, but only into H2A and H2B during G2-per1od. Resolution of acetylated H4-subspecies reveals acetate incorporation into preexisting H4, but not into newly synthesized molecules during mitosis and early S-phase. In a protam1ne competition assay histones from S-phase chromatin are released at lower protamine concentrations as compared to the lower acetylated Gi-chromatin. He demonstrate a preferential release of highly acetylated H4-subspecies at low protamine concentrations. Our results fit Into a general model of the relationship between histone acetylation and chromatin assembly. According to this model acetylation of core histones would serve as a signal for displacement of histones from nucleosomes by modulating hi stone-protein or h1stone-0...
The cell-cycle progression of germinating embryos of maize ( Z e a m a y s L.) was studied from 0... more The cell-cycle progression of germinating embryos of maize ( Z e a m a y s L.) was studied from 0 to 72 h after the start of imbibition using DNA flow cytometry on isolated nuclei, and analyses of thymidine kinase activity, histone biosynthesis and levels of proliferating cell nulcear antigen (PCNA). At the start of germination, 75% of the cells were in G1, but this population had decreased to 25% by 72 h. The concomitant increase of cells in S-phase did not occur continuously, but stepwise, indicating that during germination most of the cells enter S-phase as a partially synchronized population. Within the initial 60 h of embryo germination the cells passed through one S-phase; the start and duration of this period of replicative DNA synthesis was further substantiated by the analysis of S-phase-associated events, the biosynthesis of core histones and the enzyme activity of thymidine kinase, which both began to increase at about 12 h after the start of differentiation. Thymidine ki...
ADP-ribosylation of histones and non-histone nuclear proteins was studied in isolated nuclei duri... more ADP-ribosylation of histones and non-histone nuclear proteins was studied in isolated nuclei during the naturally synchronous cell cycle of Physarum polycephalum. Aside from ADP-ribosyltransferase (ADPRT) itself, histones and high mobility group-like proteins are the main acceptors for ADP-ribose. The majority of these ADP-ribose residues is NH2OH-labile. ADP-ribosylation of the nuclear proteins is periodic during the cell cycle with maximum incorporation in early to mid G2-phase. In activity gels two enzyme forms with Mr of 115,000 and 75,000 can be identified. Both enzyme forms are present at a constant ratio of 3:1 during the cell cycle. The higher molecular mass form cannot be converted in vitro to the low molecular mass form, excluding an artificial degradation during isolation of nuclei. The ADPRT forms were purified and separated by h.p.l.c. The low molecular mass form is inhibited by different ADPRT inhibitors to a stronger extent and is the main acceptor for auto-ADP-ribosy...
dThd bound to epoxy-activated Sepharose results in a high specific affinity adsorbent, which was ... more dThd bound to epoxy-activated Sepharose results in a high specific affinity adsorbent, which was used to purify Physarum dThd kinase 8000-fold. With this purified enzyme, it was proved that dThd kinase exists in five enzyme variants in Physarum polycephalum, the acidic forms being phosphorylated. This protein modification is discussed under the aspect of the regulation of this enzyme.
Lamin-like proteins have been identified as components of the nuclear matrix of the myxomycete Ph... more Lamin-like proteins have been identified as components of the nuclear matrix of the myxomycete Physarum polycephalum. A 67 kDa homologue was detected by immunoblotting of nuclear matrix proteins with affinity-purified anti-lamin antibodies of human autoimmune sera. A 65 kDa lamin B homologue was identified with polyclonal antibodies against turkey erythrocyte lamin B in nuclei and nuclear matrix of Physarum amoebae and plasmodia. Indirect immunofluorescence microscopy revealed that the 65 and 67 kDa proteins are localized in the nuclear lamina of plasmodial nuclei.
Interferon regulatory factor (IRF)-1 expression was surveyed in nontransformed and oncogene-trans... more Interferon regulatory factor (IRF)-1 expression was surveyed in nontransformed and oncogene-transformed mouse fibroblasts, using Western immunoblot with an IRF-1-specific antiserum, to examine possible differences resulting from cellular transformation. Ten additional proteins that reacted with the IRF-1 antibody and that underwent specific competition by peptide antigen were observed in extracts of both nontransformed and oncogene-transformed cell lines. Cross-reacting proteins were also observed in mouse macrophage extracts. Protein was captured from fibroblast nuclear extracts, using oligonucleotides representing IRF-binding sequences linked to magnetic beads. Captured proteins were eluted and analyzed by immunoblot with anti-IRF-1. Along with 43-kDa IRF-1, 4 of the 7 nuclearly located cross-reacting proteins (97, 90, 66, and 33 kDa) were found to complex with the IRF binding element. These proteins, with an epitope in common with the IRF-1 C-terminal region and IRF element DNA sequence-binding capability, may represent new members of the IRF family.
... J. 303, 723−729. Sendra, R., Rodrigo, I., Salvador, ML, &amp;amp; Franco, L. (1988) Plant... more ... J. 303, 723−729. Sendra, R., Rodrigo, I., Salvador, ML, &amp;amp; Franco, L. (1988) Plant Mol. Biol. 11, 857−866. ... Antonello Mai, Silvio Massa, Dante Rotili, Silvia Simeoni, Rino Ragno, Giorgia Botta, Angela Nebbioso, Marco Miceli, Lucia Altucci, and Gerald Brosch. ...
Hintergrund: Halsschmerzen sind eine der häufigsten Konsultationsursachen in der AM-Praxis. Bei K... more Hintergrund: Halsschmerzen sind eine der häufigsten Konsultationsursachen in der AM-Praxis. Bei Kindern sind ca. 20% und bei Erwachsenen <5% ß-hämolysierende Streptokokken der Gruppe A zu erwarten. 75% der Erwachsenen verlassen die Ordination aber mit einer Antibiotikaverordnung.[for full text, please go to the a.m. URL]
The readout of the genetic information of eukaryotic organisms is significantly regulated by modi... more The readout of the genetic information of eukaryotic organisms is significantly regulated by modifications of DNA and chromatin proteins. Chromatin alterations induce genome-wide and local changes in gene expression and affect a variety of processes in response to internal and external signals during growth, differentiation, development, in metabolic processes, diseases, and abiotic and biotic stresses. This review aims at summarizing the roles of histone H1 and the acetylation and methylation of histones in filamentous fungi and links this knowledge to the huge body of data from other systems. Filamentous fungi show a wide range of morphologies and have developed a complex network of genes that enables them to use a great variety of substrates. This fact, together with the possibility of simple and quick genetic manipulation, highlights these organisms as model systems for the investigation of gene regulation. However, little is still known about regulation at the chromatin level i...
Postsynthetic acetylation of core histones exhibits a peak during S-phase of the Physarum cell cy... more Postsynthetic acetylation of core histones exhibits a peak during S-phase of the Physarum cell cycle. The maximum H-acetate Incorporation precedes the maximum of hi stone synthesis. Acetate is incorporated into all core histones during S-phase, but only into H2A and H2B during G2-per1od. Resolution of acetylated H4-subspecies reveals acetate incorporation into preexisting H4, but not into newly synthesized molecules during mitosis and early S-phase. In a protam1ne competition assay histones from S-phase chromatin are released at lower protamine concentrations as compared to the lower acetylated Gi-chromatin. He demonstrate a preferential release of highly acetylated H4-subspecies at low protamine concentrations. Our results fit Into a general model of the relationship between histone acetylation and chromatin assembly. According to this model acetylation of core histones would serve as a signal for displacement of histones from nucleosomes by modulating hi stone-protein or h1stone-0...
The cell-cycle progression of germinating embryos of maize ( Z e a m a y s L.) was studied from 0... more The cell-cycle progression of germinating embryos of maize ( Z e a m a y s L.) was studied from 0 to 72 h after the start of imbibition using DNA flow cytometry on isolated nuclei, and analyses of thymidine kinase activity, histone biosynthesis and levels of proliferating cell nulcear antigen (PCNA). At the start of germination, 75% of the cells were in G1, but this population had decreased to 25% by 72 h. The concomitant increase of cells in S-phase did not occur continuously, but stepwise, indicating that during germination most of the cells enter S-phase as a partially synchronized population. Within the initial 60 h of embryo germination the cells passed through one S-phase; the start and duration of this period of replicative DNA synthesis was further substantiated by the analysis of S-phase-associated events, the biosynthesis of core histones and the enzyme activity of thymidine kinase, which both began to increase at about 12 h after the start of differentiation. Thymidine ki...
ADP-ribosylation of histones and non-histone nuclear proteins was studied in isolated nuclei duri... more ADP-ribosylation of histones and non-histone nuclear proteins was studied in isolated nuclei during the naturally synchronous cell cycle of Physarum polycephalum. Aside from ADP-ribosyltransferase (ADPRT) itself, histones and high mobility group-like proteins are the main acceptors for ADP-ribose. The majority of these ADP-ribose residues is NH2OH-labile. ADP-ribosylation of the nuclear proteins is periodic during the cell cycle with maximum incorporation in early to mid G2-phase. In activity gels two enzyme forms with Mr of 115,000 and 75,000 can be identified. Both enzyme forms are present at a constant ratio of 3:1 during the cell cycle. The higher molecular mass form cannot be converted in vitro to the low molecular mass form, excluding an artificial degradation during isolation of nuclei. The ADPRT forms were purified and separated by h.p.l.c. The low molecular mass form is inhibited by different ADPRT inhibitors to a stronger extent and is the main acceptor for auto-ADP-ribosy...
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