TY - JOUR A1 - Schartl, Manfred T1 - A sex chromosomal restriction-fragment-length marker linked to melanoma-determining Tu loci in Xiphophorus N2 - No abstract available KW - Physiologische Chemie Y1 - 1988 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61842 ER - TY - JOUR A1 - Riehl, R. A1 - Schartl, Manfred T1 - A Transmission Electron Microscopical and Freeze-Etch Study of Malignant-Melanoma in Fish N2 - No abstract available KW - Physiologische Chemie Y1 - 1984 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61916 ER - TY - JOUR A1 - Cavari, Benzion A1 - Hong, Yunhan A1 - Funkenstein, Bruria A1 - Moav, Boaz A1 - Schartl, Manfred T1 - All-fish gene constructs for growth hormone gene transfer in fish N2 - In order to develop all-fish expression vectors for microinjection into fertilized fish eggs, we have prepared the following cunstructs: rainbow trout metallothionein a/b and the gilthead seabream growth hormone cDNA (ptMTa-gbsGHcDNA, ptMTb-gsbGHcDNA), carp ß-actin gilthead seabream GH cDNA (pcAßgsbGHcDNA). The inducible metallothionein promoters a and b were cloned from rainbow trout, and the constitutive promoter ß-actin was isolated from carp. The metallothionein promoters were cloned by using the PCR technique. The tMTa contains 430 bp, while the tMTb contains 260 bp (Hong et al. 1992). These two promoters were introduced to pGEM-3Z containing the GH cDNA of Sparus aurata to form ptMTa-gsbGH and ptMTb-gsbGH, respectively. The carp cytoplasmic ß-actin gene was chosen as a source for isolating strong constitutive regulatory sequences. One of these regulatory sequences in pUC118 was Iigated to GH cDNA of S. aurata to form the pcAß-gsbGHcDNA. Expression of the constructs containing the metallothionein promoters was tested in fish cell culture and was found tobe induced effectively by zinc. The ptMTa gsb-GH cDNA construct was microinjected into fertilized carp eggs, and integration in the genome of carp was detected in the DNA isolated from fins at the age of two months. KW - Physiologische Chemie KW - growth hormone gene KW - all-fish genes KW - transgenic fish KW - cellline transfection KW - Sparus aurata Y1 - 1993 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61612 ER - TY - JOUR A1 - Förnzler, Dorothee A1 - Wittbrodt, Joachim A1 - Schartl, M.anfred T1 - Analysis of an esterase linked to a locus involved in the regulation of the melanoma oncogene and isolation of polymorphic marker sequences in Xiphophorus N2 - Melanoma formation in Xiphophorus hybrids is mediated by a growth factor receptor tyrosine kinase oncogene encoded by the Tu locus. In the wild-type parental fish no tumors occur due to the activity of a locus that regulates the activity of the melanoma oncogene. Molecu/ar identification of this regulatory locus (R) requires a precise physical map of the chromosomal region. Therefore we studied esterase isozymes in Xiphophorus, two of which have been previously reported to be linked to locus R. We confinn that ES 1 is a distant marker for R ( approx. 30cM), and contrary to earlier studies, we show that this isozyme is present in all species of the genus and at similar activity Ievels in all organs tested. ES4, which has also been reported to be linked to R, was found to be a misclassification of liver ES1. In an attempt to identify markersthat bridge the large distance between ESl and R, we have generated DNA probes which are highly polymorphic. They will be useful in finding Iandmarks on a physical map of the R-containing chromosomal region. KW - Physiologische Chemie KW - Xiphophorus KW - melanoma ; oncogene regulation ; esterase ; molecular marker sequences Y1 - 1991 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61726 ER - TY - JOUR A1 - Malitschek, Barbara A1 - Wittbrodt, Joachim A1 - Fischer, Petra A1 - Lammers, Reiner A1 - Ullrich, Axel A1 - Schartl, Manfred T1 - Autocrine stimulation of the Xmrk receptor tyrosine kinase in Xiphophorus melanoma cells and identification of a source for the physiological ligand N2 - The melanoma·inducing gene of Xiphophorus fish encodes the Xmrk receptor tyrosine kinase. U sing a highly specific antiserum p~oduced against the recombinant receptor expressed with a baculovirus, it is shown that Xmrk is the most abundant phosphotyrosine protein in fish melanoma and thus highly activated in the tumors. Studies on a melanoma cellline revealed that these cells produce an activity that considerably stimulates receptor autophosphorylation. The stimulating activity induces receptor down-regulation and can be depleted from the melanoma cellsupernatant by the immobilized recombinant receptor protein. The fish melanoma cells can thus be considered autocrine tumor cells providing a source for future purification and characterization of the Xmrk ligand. KW - Physiologische Chemie Y1 - 1994 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61551 ER - TY - JOUR A1 - Götz, Rudolf A1 - Raulf, Friedrich A1 - Schartl, Manfrad T1 - Brain-derived neurotrophic factor is more highly conserved in structure and function than nerve growth factor during vertebrate evolution N2 - Mammalian nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are members of a protein family with perfectly conserved domains arranged around the cysteine residues thought to stabilize an invariant three-dimensional scaffold in addition to distinct sequence motifs that convey different neuronal functions. To study their structural and functional conservation during evolution, we have compared NGF and BDNF from a lower vertebrate, the teleost fi.sh Xiphophorus, with the mammalian homlogues. Genomic clones encoding fish NGF and BDNF were isolated by cross-hybridization using probes from the cloned mammalian factors. Fish NGF and BDNF were expressed by means of recombinant vaccinia viruses, purified, and their neuronal survival specificities for different classes of neurons were found to mirror those of the mammalian factors. The half-maximal survival concentration for chick sensory neurons was 60 pg/ml for both fish and mammalian purifi.ed recombinant BDNF. However, the activity ofrecombinant fish NGF on both chick sensory and sympathetic neurons was 6 ng,lml, 75-fold lower than that of mouse NGF. The different functional conservation of NGF and BDNF is also reflected in their structures. The DNA-deduced amino acid sequences of processed mature fish NGF and BDNF showed, compared to mouse, 63% and 90% identity, respectively, indicating that NGF bad reached an optimized structure later than BDNF. The retrograde extrapolation of these data indicates that NGF and BDNF evolved at strikingly different rates ftom a common ancestral gene about 600 million years ago. By RNA gel blot anaJysis NGF mRNA was detected during late embryonie development; BDNF was present in adult brain. KW - Physiologische Chemie KW - Brain-derived neurotrophic factor KW - Nerve growth factor KW - Fish KW - Recombinant protein expression KW - Neuronal survival Y1 - 1992 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61703 ER - TY - JOUR A1 - Schartl, Manfred A1 - Barnekow, A. T1 - Cellular src gene product detected in the freshwater sponge Spongilla lacustris N2 - No abstract available KW - Physiologische Chemie Y1 - 1984 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61904 ER - TY - JOUR A1 - Barnekow, A. A1 - Schartl, Manfred T1 - Comparative studies on the src proto-oncogene and its gene product pp60\(^{c-src}\) in normal and neoplastic tissues of lower vertebrates N2 - No abstract available KW - Physiologische Chemie Y1 - 1987 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61869 ER - TY - JOUR A1 - Riehl, R. A1 - Schartl, Manfred A1 - Kollinger, G. T1 - Comparative studies on the ultrastructure of malignant melanoma in fish and human by freeze-etching and transmission electron microscopy N2 - Malignant melanomas (MM) in the fish Xiphophorus and in humans were studied both by transmission electron microscopy (TEM) and freeze-etching (FE). In both fish and human melanomas the cells show interdigitations of the,plasma membranes. The nuclei are large and lobulated and have many nuclear pores. Melanosomes are abundant and melanosome complexes ("compound melanosomes") occur regularly. Pinocytotic vesicles could be demonstrated in fish and human melanomas showing iocal differences in frequency and distribution patterns in the tumor. lntercellular junctions are lacking in MM cells from fish and humans. The FE technique showed considerable advantages in demonstrating membrane-surface peculiarities such as nuclear pores or pinocytotic vesicles. The FE replicas of fish melanomas are like those of humans. These findings may support the hypothesis that melanoma in fish and humans reflect the same biological phenomenon. KW - Physiologische Chemie KW - Malignant melanoma KW - Fish KW - Human KW - Freeze-etching KW - Transmission electron microscopy Y1 - 1984 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61920 ER - TY - JOUR A1 - Schartl, Manfred A1 - Barnekow, A. A1 - Bauer, H. A1 - Anders, F. T1 - Correlations of inheritance and expression between a tumor gene and the cellular homolog of the Rous sarcoma virus-transforming gene in Xiphophorus N2 - No abstract available KW - Physiologische Chemie Y1 - 1982 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61937 ER - TY - JOUR A1 - Birkmayer, G. D. A1 - Sebald, Walter A1 - Bücher, Th. T1 - Cytochrom-Oxydase und ein an diese assoziiertes, markiertes Protein aus 14C-markierten Mitochondrien von Neurospora crassa T1 - Cytochrome oxidase and associated with it a labelled protein from 14C-labelled mitochondria of Neurospora crassa N2 - no abstract available KW - Physiologische Chemie Y1 - 1969 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-82135 ER - TY - JOUR A1 - Hong, Yunhan A1 - Winkler, Christoph A1 - Brem, Gottfried A1 - Schartl, Manfred T1 - Development of a heavy metal-inducible fish-specific expression vector for gene transfer in vitro and in vivo N2 - The promoter of the rainbow trout metallothionein B gene ( tMTb) was isolated from genomic DNA by the polymerase chain reaction (PCR ), fused to the bacterial chloramphenicol acetyltransferase (CAT) genein an expression vector, and functionally analyzed in one human cellline and four fish celllines. This promoter exhibited an extremely low basal expression in all celllines and was zincand cadmium-inducible except in the fish melanoma cell line where the promoter was completely inactive. The metal-induced expression patterns were cellline-specific. In general the fish promoter was more species- and cell type-specific than its human counterpart. In a transient assay it was functional in developing embryos of the medaka ( Oryzias /atipes). These properties make this promoter suitable for inducible, tissue-specific expression of transgenes and for in vivo studies of gene function and regulation. KW - Physiologische Chemie Y1 - 1993 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61666 ER - TY - THES A1 - Heinecke, Kai T1 - Die Dynamik der primären Erkennungsschritte von BMP-Rezeptoren T1 - Dynamics of the primary recognition steps of BMP receptors N2 - Bone Morphogenetic Proteins (BMPs) bilden zusammen mit den Activinen, Growth and Differentiation Factors (GDFs) und Transforming Growth Factor β (TGF-β) die Transforming Growth Factor β-Superfamilie von sekretierten Signalproteinen. Sie spielen eine wichtige Rolle in der Entwicklung, Erhaltung und Regeneration von Geweben und Organen. Die Signalvermittlung dieser Proteine erfolgt durch die Bindung von zwei verschiedenen Typen von Serin-/Threonin-Kinaserezeptoren, die als Typ-I- und Typ-II-Rezeptoren bezeichnet werden. Im ersten Schritt erfolgt die Bindung an den hochaffinen Rezeptor (im Fall von BMP-2 der Typ-I-Rezeptor), im nächsten Schritt wird der niederaffine Rezeptor in den Komplex rekrutiert. Bis heute sind lediglich sieben Typ-I- und fünf Typ-II-Rezeptoren bekannt, was auf eine Promiskuität in der Liganden-Rezeptor-Interaktion schließen lässt. Die Architektur beider Rezeptorsubtypen ist dabei relativ ähnlich. Beide bestehen aus einer ligandenbindenden extrazellulären Domäne, einer Transmembrandomäne sowie einer intrazellulären Kinasedomäne. Eine nacheinander ablaufende Transphosphorylierung der intrazellulären Domänen führt zu einer Phosphorylierung von SMAD-Proteinen, die dann als nachgeschaltete Vermittler fungieren und die Transkription regulierter Gene auslösen. Im Hauptteil dieser Arbeit wurden die initialen Schritte der Rezeptorkomplexformierung sowie die Mobilität der Rezeptoren mit Hilfe von fluoreszenzmikroskopischen Methoden untersucht. Dabei konnte festgestellt werden, dass für die Bildung eines Signalkomplexes eine bestimmte Schwellenkonzentration des Liganden nötig ist und dass der Mechanismus nach einem Alles-oder-Nichts-Prinzip wie ein Schalter funktioniert. Außerdem konnten Unterschiede in der Nutzung der gleichen Rezeptoren durch verschiedene Liganden festgestellt werden. Die anderen Teile der Arbeit befassen sich mit der Funktionalität der verschiedenen Rezeptordomänen in der Signalübermittlung, der Analyse von hoch- und niederaffinen Ligandenbindestellen auf ganzen Zellen sowie dem Einfluss des SMAD- und des MAPK-Signalwegs auf die Induktion der Alkalischen Phosphatase. Dabei konnte gezeigt werden, dass die Art der SMAD-Phosphorylierung allein vom Typ der Kinasedomäne abhängig ist, dass auf einer Zelle verschiedene Rezeptorpopulationen existieren, welche von unterschiedlichen Ligandenkonzentrationen angesprochen werden, und dass die Induktion der Alkalischen Phosphatase stark vom zeitlichen Verlauf der SMAD- und MAPK-Aktivierung abhängig ist. N2 - Bone Morphogenetic Proteins (BMPs), together with Activins, Growth and Differentiation Factors (GDFs) and Transforming Growth Factor β (TGFβ), are secreted signalling proteins that belong to the Transforming Growth Factor β superfamily. They play an important role in regulating the development, maintenance and regeneration of tissues and organs. Signalling of TGFβ superfamily members occurs by binding to two types of serine-/threonine kinase receptors termed type I and type II. First, the high affinity receptor (in case of BMP2 the type I receptor) is bound, and then the low affinity receptor is recruited into the signalling complex. The fact that there are only seven type-I and five type-II receptors are known implies a limited promiscuity in ligand-receptor interaction. The architecture of both receptor subtypes is quite similar, with a small extracellular ligand-binding domain, a single transmembrane domain and an intracellular kinase domain. Subsequent transphosphorylation of the intracellular receptor domains leads to phosphorylation of SMAD proteins, which then act as downstream mediators and activate gene transcription. The main part of this work was to analyze the initial steps in receptor complex formation and the mobility of TGFβ-superfamily receptors with fluorescence microscopy techniques. It could be shown that complex formation requires a certain ligand threshold concentration and shows an all-or-nothing switch-like behaviour. Furthermore, differences between different ligands using the same receptors could be visualized. The other parts of this work deal with the functionality of the different receptor domains in signal transduction, the analysis of high- and low-affinity binding sites on whole cells and the influence of the SMAD- and MAPK-pathways on alkaline phosphatase induction. It could be shown that the type of SMAD phosphorylation ist solely dependent on the type of the kinase domain, that there exist different receptor populations on a cell that are addressed by different ligand concentrations and that alkaline phosphatase induction is highly dependent on the time course of SMAD- and MAPK-pathway activation. KW - Knochen-Morphogenese-Proteine KW - Wirkstoff-Rezeptor-Bindung KW - Signaltransduktion KW - Signalkomplex KW - Rezeptormobilität KW - Rezeptor-Liganden-Interaktion KW - Physiologische Chemie KW - Molekulare Biophysik KW - Molekularbiologie KW - Signaling complex KW - receptor mobility KW - receptor-ligand-interaction Y1 - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-49257 ER - TY - JOUR A1 - Mäueler, Winfried A1 - Schartl, Angelika A1 - Schartl, Manfred T1 - Different expression patterns of oncogenes and proto-oncogenes in hereditary and carcinogen-induced tumors of Xiphophorus N2 - Hereditary melanoma in Xiphophorus hybrids canying the melanoma·induclng Tu-Sd locus is caused by transcriptional activation of the Xmrk gene that resides at the Tu·Sd locus and encodes a novel member of receptor tyrosine kinases (RTK). ln this study, a total of 17 hereditary melanomas from various hybrid genotypes harbouring 7 different Tu alleles were also found to aver-express the correspondlng Xmrlc alleles. The Ievei of over-expression correlated with the degree of malignancy of the melanoma. ln addition, Xsrc expression was high ln many malignant melanomas. Expression pattems and Ieveis of the Xiphophorus EGF-receptor gene (Xerb B), the c-myc (Xmyc), and the PDGF (Xsls) gene(s) were not intriguing. Transcription of the ras gene(s) may be correlated to secondary events of melanoma progression. Expression pattems of Xfms, the Xiphophorus CSF-1 receptor homologue, can be explained by different contents of infiltrating macrophages in the tumors. ln carcinogen-induced tumors includlng one melanoma no significant expression of the Xmrk oncogene could be detected. Xsrc expression, however, was strikingly high. This indicates that activation of oncogenes other than Xmrk ls instrumental in tumorigenesls of neoplasia of non-hereditary origin. KW - Physiologische Chemie Y1 - 1993 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61592 ER - TY - JOUR A1 - Schartl, Manfred A1 - Barnekow, A. T1 - Differential expression of the cellular src gene during vertebrate development N2 - No abstract available KW - Physiologische Chemie Y1 - 1984 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61893 ER - TY - JOUR A1 - Nanda, Indrajit A1 - Schartl, Manfred A1 - Feichtinger, Wolfgang A1 - Epplen, Jörg T. A1 - Schmid, Michael T1 - Early stages of sex chromosome differentiation in fish as analysed by simple repetitive DNA sequences N2 - Animal sex chromosome evolution has started on different occasions with a homologous pair of autosomes leading to morphologically differentiated gonosomes. In contrast to other vertebrate classes, among fishes cytologically dernonstrahle sex chromosomes are rare. In reptiles, certain motifs of simple tandemly repeated DNA sequences like (gata)\(_n\)/(gaca)\(_m\) are associated with the constitutive heterochromatin of sex chromosomes. In this study a panel of simple repetitive sequence probes was hybridized to restriction enzyme digested genomic DNA of poeciliid fishes. Apparent male heterogamety previously established by genetic experiments in Poecilia reticulata (guppy) was correlated with male-specific hybridization using the (GACA)\(_4\) probe. The (GATA)\(_4\) oligonucleotide identifies certain male guppies by a Y chromosomal polymorphism in the outbred population. In cantrast none of the genetically defined heterogametic situations in Xiphophorus could be verified consistently using the collection of simple repetitive sequence probes. Only individuals from particular populations produced sex-specific patterns of hybridization with (GATA)\(_4\). Additional poeciliid species (P. sphenops, P. velifera) harbour different sex-specifically organized simple repeat motifs. The observed sex-specific hybridization patterns were substantiated by banding analyses of the karyotypes and by in situ hybridization using the (GACA)\(_4\) probe. KW - Physiologische Chemie Y1 - 1992 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61715 ER - TY - JOUR A1 - Schartl, Manfred A1 - Schmidt, C. R. A1 - Anders, A. A1 - Barnekow, A. T1 - Elevated expression of the cellular src gene in tumors of differing etiologies in Xiphophorus N2 - No abstract available KW - Physiologische Chemie Y1 - 1985 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61889 ER - TY - JOUR A1 - Raulf, F. A1 - Robertson, S. M. A1 - Schartl, Manfred T1 - Evolution of the neuron-specific alternative splicing product of the c-src proto-oncogene N2 - The observation of a slower migrating form of pp6oc-src in neural tissue of chicken and mouse has recently been shown to be due to an alternative transcript form of tbe c-src gene (Martinez et al.: Science 237:411-415, 1987; Levy et al.: Mol Cell Bio17:4142- 4145, 1987). An insertion of 18 basepairs between exons 3 and 4, presumed to be due to alternative splicing of a mini-exon, gives rise to six amino acid residues not found in the non-neuronal (termed flbroblastic) form of pp60\(^{c-src}\). Wehave addressed the question of the evolutionary origin of the c-src neuronal insert · and its functional signiflcance regarding neural-speciflc expression of the c-src gene. To this end we have investigated whether the c-src gene of a lower verlebrate (the teleost fish Xiphophorus) gives rise to a neural-specific transcript in an analogous manner. We could show that the fish c-src gene does encode for a "fibroblastic" and a "neuronal" form of transcript and that the neuronal transcript does indeed arise by way of alternative splicing of a mini-exon. The miniexon is also 18 basepairs long and we could demoostrate directly that this exon lies within the intron separating exons 3 and 4. For comparative purposes we have examined whether the fish c-yes gene, the member of the src gene family most closely related to c-src, also encodes a neural tissue-specific transcript. No evidence for a second transcript form in brain was obtained. This result suggests that the mini-exon arose within the c-src gene lineage sometime between the srclyes gene duplication event and the divergence of the evolutionary lineage giving rise to the teleost fish. Published genomic sequence of src-related genes in Drosophila and our own results with Hydra demoostrate no intron in these species at the analogous location, consistent with first appearance of this mini-exon sometime between 550 and 400 million years ago. KW - Physiologische Chemie KW - Xiphophorus KW - teleost flsh KW - polymerase KW - chain reaction KW - RT -PCR KW - mini-exon KW - pp6oc-src Y1 - 1989 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61796 ER - TY - JOUR A1 - Schartl, A. A1 - Dimitrijevic, N. A1 - Schartl, Manfred T1 - Evolutionary origin and molecular biology of the melanoma-inducing oncogene of Xiphophorus N2 - No abstract available KW - Physiologische Chemie Y1 - 1994 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61954 ER - TY - JOUR A1 - Barnekow, A. A1 - Paul, E. A1 - Schartl, Manfred T1 - Expression of the c-src protooncogene in human skin tumors N2 - No abstract available KW - Physiologische Chemie Y1 - 1987 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-61870 ER -