TY - JOUR A1 - Wallschläger, Hans-Dieter A1 - Beier, Wolfgang A1 - Burkart, Michael A1 - Mrzljak, Jadranka A1 - Oehlschläger, Susanne A1 - Wanner, Manfred T1 - Ökologische Datenerfassung für Naturschutzbewertung und Monitoring im Offenland Y1 - 2004 SN - 3-540-22449-1 ER - TY - JOUR A1 - Seitz, Birgit A1 - Ristow, Michael A1 - Klemm, Gunther A1 - Rätzel, Stefan A1 - Schulze, Gerhart A1 - Hoffmann, Maik T1 - Zur Verbreitung der Wildrosen und verwilderten Kulturrosen in Berlin und Brandenburg Y1 - 2004 SN - 0724-3111 - ER - TY - JOUR A1 - Albrecht, Tanja A1 - Haebel, Sophie A1 - Koch, Anke A1 - Krause, Ulrike A1 - Eckermann, Nora A1 - Steup, Martin T1 - Yeast glycogenin (Glg2p) produced in Escherichia coli is simultaneously glucosylated at two vicinal tyrosin residues but results in a reduced bacterial glycogen accumulation N2 - Saccharomyces cerevisiae possesses two glycogenin isoforms (designated as Glg1p and Glg2p) that both contain a conserved tyrosine residue, Tyr232. However, Glg2p possesses an additional tyrosine residue, Tyr230 and therefore two potential autoglucosylation sites. Glucosylation of Glg2p was studied using both matrix-assisted laser desorption ionization and electrospray quadrupole time of flight mass spectrometry. Glg2p, carrying a C-terminal (His(6)) tag, was produced in Escherichia coli and purified. By tryptic digestion and reversed phase chromatography a peptide (residues 219-246 of the complete Glg2p sequence) was isolated that contained 4-25 glucosyl residues. Following incubation of Glg2p with UDPglucose, more than 36 glucosyl residues were covalently bound to this peptide. Using a combination of cyanogen bromide cleavage of the protein backbone, enzymatic hydrolysis of glycosidic bonds and reversed phase chromatography, mono- and diglucosylated peptides having the sequence PNYGYQSSPAM were generated. MS/MS spectra revealed that glucosyl residues were attached to both Tyr232 and Tyr230 within the same peptide. The formation of the highly glucosylated eukaryotic Glg2p did not favour the bacterial glycogen accumulation. Under various experimental conditions Glg2p-producing cells accumulated approximately 30% less glycogen than a control transformed with a Glg2p lacking plasmid. The size distribution of the glycogen and extractable activities of several glycogen-related enzymes were essentially unchanged. As revealed by high performance anion exchange chromatography, the intracellular maltooligosaccharide pattern of the bacterial cells expressing the functional eukaryotic transgene was significantly altered. Thus, the eukaryotic glycogenin appears to be incompatible with the bacterial initiation of glycogen biosynthesis Y1 - 2004 ER - TY - JOUR A1 - Pacholsky, Dirk A1 - Vakeel, Padmanabhan A1 - Himmel, Mirko A1 - Lowe, T. A1 - Stradal, T. A1 - Rottner, K. A1 - Fürst, Dieter Oswald A1 - vanderVen, Peter F. M. T1 - Xin repeats define a novel actin-binding motif N2 - Xin is a protein that is expressed during early developmental stages of cardiac and skeletal muscles. Immunolocalization studies indicated a peripheral localization in embryonic mouse heart, where Xin localizes with beta- catenin and N-cadherin. In adult tissues, Xin is found primarily in the intercalated discs of cardiomyocytes and the myotendinous junctions of skeletal muscle cells, both specialized attachment sites of the myofibrillar ends to the sarcolemma. A large part of the Xin protein consists of unique 16 amino acid repeats with unknown function. We have investigated the characteristics of the Xin repeats by transfection experiments and actin-binding assays and ascertained that, upon expression in cultured cells, these repeats bind to and stabilize the actin-based cytoskeleton. In vitro co- sedimentation assays with skeletal muscle actin indicated that they not only directly bind actin filaments, but also have the capability of arranging microfilaments into networks that sediment upon low-speed centrifugation. Very similar repeats were also found in Xin-repeat protein 2' (XIRP2), a novel protein that seems to be expressed mainly in striated muscles. Human XIRP2 contains 28 Xin repeats with properties identical to those of Xin. We conclude that the Xin repeats define a novel, repetitive actin-binding motif present in at least two different muscle proteins. These Xin- repeat proteins therefore constitute the first two members of a novel family of actin-binding proteins Y1 - 2004 SN - 0021-9533 ER - TY - JOUR A1 - Kummer, Volker T1 - Vom Seidenbau in Krausnick Y1 - 2004 ER - TY - JOUR A1 - Boese, Stefan H. A1 - Gray, Michael A. A1 - Simmons, N. L. T1 - Volume and non-volume activated anion conductances and their interactions in the renal IMCD Y1 - 2004 SN - 0-387- 23299-0 ER - TY - JOUR A1 - Weithoff, Guntram T1 - Vertical niche separation of two consumers (Rotatoria) in an extreme habitat N2 - Herbivore populations are commonly restricted by resource limitation, by predation or a combination of the two. Food supplement experiments are suitable for investigating the extent of food limitation at any given time. The main part of this study was performed in an extremely acidic lake (pH 2.7) where the food web consists of only a few components and potential food sources for herbivores are restricted to two flagellates. Life table experiments proved that Chlamydomonas was a suitable food source whereas Ochromonas was an unsuitable food source. The two flagellates and the two rotifers exhibit a pronounced vertical distribution pattern. In this study, a series of food supplement experiments were performed in order to: (1) quantify and compare potential resource limitation of two primary consumers (Cephalodella hoodi and Elosa worallii, Rotatoria) over time, (2) compare their response at different temperatures, (3) evaluate the effect of having an unsuitable food source alongside a valuable one, (4) estimate the effect of predation on rotifers by Heliozoa, and (5) compare the results with those from other acidic lakes. Additionally, the spatio- temporal population dynamics of both species were observed. The field data confirmed a vertical separation of the two species with E. worallii dominating in the upper water layers, and C. hoodi in the deeper, cooler water layers. The results from the food supplement experiments in which Chlamydomonas served as the supplemented suitable food source showed that the two rotifers were food limited in the epilimnion throughout the season to different extents, with Cephalodella being more severely food limited than Elosa. The experiments at different temperatures provided evidence that Elosa had a higher optimum temperature for growth than Cephalodella. When the unsuitable food algae Ochromonas was added alongside the suitable food source Chlamydomonas, C. hoodi was unaffected but E. worallii was negatively affected. Predation of Heliozoa on rotifers was observed but the total effect on the rotifer dynamics is probably low. The comparison with other lakes showed that resource limitation also occurred in one other lake, although to a lesser extent. Overall, the vertical separation of the two rotifers could be explained by both their differential extent of resource limitation and differential response to temperature. Y1 - 2004 ER - TY - JOUR A1 - Hilson, Pierre A1 - Allemeersch, Joke A1 - Altmann, Thomas A1 - Aubourg, Sebastien A1 - Avon, Alexandra A1 - Beynon, Jim A1 - Bhalerao, Rishikesh P. A1 - Bitton, Frederique A1 - Caboche, Michel A1 - Cannoot, Bernard A1 - Chardakov, Vasil A1 - Cognet-Holliger, Cecile A1 - Colot, Vincent A1 - Crowe, Mark A1 - Darimont, Caroline A1 - Durinck, Steffen A1 - Eickhoff, Holger A1 - deLongevialle, Andeol Falcon A1 - Farmer, Edward E. A1 - Grant, Murray A1 - Kuiper, Martin T. R. A1 - Lehrach, Hans A1 - Leon, Celine A1 - Leyva, Antonio A1 - Lundeberg, Joakim A1 - Lurin, Claire A1 - Moreau, Yves T1 - Versatile gene-specific sequence tags for arabidopsis functional genomics : transcript profiling and reserve genetics applications N2 - Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics Y1 - 2004 ER - TY - GEN A1 - Köchy, Martin A1 - Wilson, Scott D. T1 - Variation in nitrogen deposition and available soil nitrogen in a forest–grassland ecotone in Canada N2 - Regional variation in nitrogen (N) deposition increases plant productivity and decreases species diversity, but landscape- or local-scale influences on N deposition are less well-known. Using ion-exchange resin, we measured variation of N deposition and soil N availability within Elk Island National Park in the ecotone between grassland and boreal forest in western Canada. The park receives regionally high amounts of atmospheric N deposition (22 kg ha⁻¹ yr⁻¹). N deposition was on average higher ton clayrich luvisols than on brunisols, and areas burned 1 – 15 years previously received more atmospheric N than unburned sites. We suggest that the effects of previous fires and soil type on deposition rate act through differences in canopy structure. The magnitude of these effects varied with the presence of ungulate grazers (bison, moose, elk) and vegetation type (forest, shrubland, grassland). Available soil N (ammonium and nitrate) was higher in burned than unburned sites in the absence of grazing, suggesting an effect of deposition. On grazed sites, differences between fire treatments were small, presumably because the removal of biomass by grazers reduced the effect of fire. Aspen invades native grassland in this region, and our results suggest that fire without grazing might reinforce the expansion of forest into grassland facilitated by N deposition. KW - fire KW - grazing KW - vegetation type KW - soil type KW - shrubland KW - forest invasion KW - subboreal KW - aspen parkland KW - Canada Y1 - 2004 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-5768 ER - TY - THES A1 - Usadel, Björn T1 - Untersuchungen zur Biosynthese der pflanzlichen Zellwand = [Identification and characterization of genes involved in plant cell wall synthesis] T1 - Untersuchungen zur Biosynthese der pflanzlichen Zellwand N2 - Even though the structure of the plant cell wall is by and large quite well characterized, its synthesis and regulation remains largely obscure. However, it is accepted that the building blocks of the polysaccharidic part of the plant cell wall are nucleotide sugars. Thus to gain more insight into the cell wall biosynthesis, in the first part of this thesis, plant genes possibly involved in the nucleotide sugar interconversion pathway were identified using a bioinformatics approach and characterized in plants, mainly in Arabidopsis. For the computational identification profile hidden markov models were extracted from the Pfam and TIGR databases. Mainly with these, plant genes were identified facilitating the “hmmer” program. Several gene families were identified and three were further characterized, the UDP-rhamnose synthase (RHM), UDP-glucuronic acid epimerase (GAE) and the myo-inositol oxygenase (MIOX) families. For the three-membered RHM family relative ubiquitous expression was shown using variuos methods. For one of these genes, RHM2, T-DNA lines could be obtained. Moreover, the transcription of the whole family was downregulated facilitating an RNAi approach. In both cases a alteration of cell wall typic polysaccharides and developmental changes could be shown. In the case of the rhm2 mutant these were restricted to the seed or the seed mucilage, whereas the RNAi plants showed profound changes in the whole plant. In the case of the six-membered GAE family, the gene expressed to the highest level (GAE6) was cloned, expressed heterologously and its function was characterized. Thus, it could be shown that GAE6 encodes for an enzyme responsible for the conversion of UDP-glucuronic acid to UDP-galacturonic acid. However, a change in transcript level of variuos GAE family members achieved by T-DNA insertions (gae2, gae5, gae6), overexpression (GAE6) or an RNAi approach, targeting the whole family, did not reveal any robust changes in the cell wall. Contrary to the other two families the MIOX gene family had to be identified using a BLAST based approach due to the lack of enough suitable candidate genes for building a hidden markov model. An initial bioinformatic characterization was performed which will lead to further insights into this pathway. In total it was possible to identify the two gene families which are involved in the synthesis of the two pectin backbone sugars galacturonic acid and rhamnose. Moreover with the identification of the MIOX genes a genefamily, important for the supply of nucleotide sugar precursors was identified. In a second part of this thesis publicly available microarray datasets were analyzed with respect to co-responsive behavior of transcripts on a global basis using nearly 10,000 genes. The data has been made available to the community in form of a database providing additional statistical and visualization tools (http://csbdb.mpimp-golm.mpg.de). Using the framework of the database to identify nucleotide sugar converting genes indicated that co-response might be used for identification of novel genes involved in cell wall synthesis based on already known genes. N2 - Obwohl der Aufbau der pflanzlichen Zellwand im Großen und Ganzen relativ gut charakterisiert ist, ist relativ wenig über ihre Synthese bekannt. Allgemein akzeptiert ist jedoch, dass die Nukleotidzucker die Vorstufe für den polysaccharidären Teil der Zellwand stellen. Im Rahmen der vorliegenden Arbeit wurden neue Kandidatengene für die Zellwandbiosynthese mittels bioinformatorischer Analysen ermittelt und deren Rolle in Pflanzen, hauptsächlich Arabidopsis thaliana untersucht. Zur Identifizierung von Arabidopsis thaliana Kandidatengenen des Nukleotidzucker-Stoffwechselweges wurden „hidden Markov Modelle“ für Gene desselben aus den Datenbanken Pfam und TIGR extrahiert. Unter anderem wurden diese dann unter Zuhilfenahme des Programms hmmer zur Identifikation von pflanzlichen Genen benutzt. Es wurden einige Genfamilien identifiziert und drei von diesen wurden weiter charakterisiert. Hierbei handelte sich um eine UDP-Rhamnose Synthase Familie (RHM), eine UDP-Glucuronsäurepimerase Familie (GAE) und eine myo-Inositol Oxygenase Familie (MIOX). Für die RHM Kandidatengenfamilie, mit drei Mitgliedern, wurde die relativ ubiquitäre Expression aller Gene mittels verschiedener Methoden gezeigt und für eines der Gene, RHM2, konnten T-DNA Linien bezogen werden. Außerdem wurde die Transkription der gesamten Familie mittels eines RNAi Konstruktes herunter geregelt. In beiden Fällen konnte eine Veränderung von zellwandtypischen Polysacchariden sowie schwere Entwicklungsstörungen gezeigt werden. Diese waren bei der rhm2 Funktionsverlustpflanze auf den Samenschleim bzw. den Samen reduziert, bei den RNAi Pflanzen hingegen war die gesamte Pflanze betroffen. Im Falle der zweiten Kandidatengenfamilie, GAE, wurde das höchst-exprimierte Gen (GAE6) kloniert, heterolog exprimiert und die Funktion charakterisiert. So konnte gezeigt werden, dass GAE6 für ein Enzym kodiert, welches UDP-Glukuronsäure in UDP-Galakturonsäure wandelt. Allerdings zeigten Pflanzen mit veränderter Transkriptmenge, erreicht durch T-DNA Insertionen (gae2, gae5, gae6), Überexpression (GAE6) oder RNAi , keine robuste Veränderung der Zellwand. Die letzte betrachtete Kandiatengenfamilie myo-Inositol Oxygenase wurde im Gegensatz zu den beiden anderen Familien, durch eine BLAST Suche gefunden, da zur Zeit der Durchführung noch zu wenig myo-inositol Oxygenasen bekannt waren, um daraus „hidden Markov Modelle“ abzuleiten. Dennoch konnten erste bioinformatorische Analysen zu dieser Genfamilie gemacht werden. Insgesamt gesehen wurden in diesem Teil der Arbeit die beiden Genfamilien identifiziert und charkterisiert, die bei der Synthese der beiden Pektinrückgradzucker Rhamnose und Galakturonsäure die tragende Rolle spielen. Weiterhin wurde mit der Identifizierung der MIOX Genfamilie, eine Genfamilie identifiziert, die wichtige Vorstufen in der Synthese der Nukleotidzucker liefert. In einem zweiten Teil der Arbeit wurden öffentlich zugängliche Mikroarray-Daten durch ihr Gleich -oder Ungleichverhalten charakterisiert. Dieses erfolgte auf globaler Ebene für zunächst fast 10.000 Gene. Die Daten wurden in Form einer allgemein zugänglichen Datenbank der Allgemeinheit zur Verfügung gestellt (http://csbdb.mpimp-golm.mpg.de). Eine Anwendung der Methode auf Gene des Nukleotidzuckerstoffwechsels, deutet darauf hin, dass so neue Kandiatengene, die bei der Zellwandsynthese eine Rolle spielen, von bereits bekannten Genen abgeleitet werden können. T2 - Identification and Characterization of Genes Involved in Plant Cell Wall Synthesis KW - Zellwand KW - Rhamnose KW - Galacturonsäure KW - Pektinsäure KW - Pektine KW - Inosite KW - Korrelationsanalyse KW - plant cell wall biosynthesis KW - udp-rhamnose KW - udp-galacturonic acid KW - correlation networks Y1 - 2004 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-2947 ER -