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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The degradation of aromatic compounds follows different biochemical principles in aerobic and anaerobic microorganisms. While aerobes dearomatize and cleave the aromatic ring by oxygenases, facultative anaerobes utilize an ATP-dependent ring reductase for the dearomatization of the activated key intermediate benzoyl-coenzyme A (CoA). In this work, the aromatic metabolism was studied in the obligately anaerobic model organism Geobacter metallireducens. The gene coding for a putative carboxylic acid-CoA ligase was heterologously overexpressed and the gene product was characterized as a highly specific benzoate-CoA ligase catalysing the initial step of benzoate metabolism. However, no evidence for the presence of an ATP-dependent benzoyl-CoA reductase as observed in facultative anaerobes was obtained. In a proteomic approach benzoate-induced proteins were identified; the corresponding genes are organized in two clusters comprising 44 genes. Induction of representative genes during growth on benzoate was confirmed by reverse transcription polymerase chain reaction. The results obtained suggest that benzoate is activated to benzoyl-CoA, which is then reductively dearomatized to cyclohexa-1,5-diene-1-carbonyl-CoA, followed by β-oxidation reactions to acetyl-CoA units, as in facultatively anaerobic bacteria. However, in G. metallireducens the process of reductive benzene ring dearomatization appears to be catalysed by a set of completely different protein components comprising putative molybdenum and selenocysteine containing enzymes.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Planta 208 (1999), S. 301-309 
    ISSN: 1432-2048
    Keywords: Key words: Cytokinin ; Developmental mutant ; Expressed sequence tag ; Gene disruption ; Homologous recombination ; Moss ; Photoreceptor ; Physcomitrella
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Key words: AMP-isopentenyl ; phosphotransferase ; Auxin ; Chloroplast division ; Cell differentiation ; Cytokinin - Physcomitrella
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Development of Physcomitrella patens (Hedw.) B.S.G. starts with a filamentous protonema growing by apical cell division. As a developmental switch, some subapical cells produce three-faced apical cells, the so-called buds, which grow to form leafy shoots, the gametophores. Application of cytokinins enhances bud formation but no subsequent gametophore development in several mosses. We used the ipt gene of Agrobacterium tumefaciens, encoding a protein which catalyzes the rate-limiting step in cytokinin biosynthesis, to transform two developmental Physcomitrella mutants. One mutant (P24) was defective in budding (bud) and thus did not produce three-faced cells, while the other one (PC22) was a double mutant, defective in plastid division (pdi), thus possessing at the most one giant chloroplast per cell, and in gametophore development (gad), resulting in malformed buds which could not differentiate into leafy gametophores. Expression of the ipt gene rescued the mutations in budding and in plastid division but not the one in gametophore development. By mutant rescue we provide evidence for a distinct physiological difference between externally applied and internally produced cytokinins. Levels of immunoreactive cytokinins and indole-3-acetic acid were determined in tissues and in culture media of the wild-type moss, both mutants and four of their stable ipt transformants. Isopentenyl-type cytokinins were the most abundant cytokinins in Physcomitrella, whereas zeatin-type cytokinins, the major native cytokinins of higher plants, were not detectable. Cytokinin as well as auxin levels were enhanced in ipt transgenics, demonstrating a cross-talk between both metabolic pathways. In all genotypes, most of the cytokinin and auxin was found extracellularly. These extracellular pools may be involved in hormone transport in the non-vascular mosses. We suggest that both mutants are defective in signal-transduction rather than in cytokinin metabolism.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 20 (1991), S. 319-329 
    ISSN: 1432-0983
    Keywords: Lower plant mitochondrial DNA ; Cytochrome oxidase subunit III gene ; Moss (Physcomitrella patens)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The first mitochondrial-encoded gene of an archeogoniate has been identified, cloned and sequenced. The cytochrome oxidase III gene (cox3) of the moss Physcomitrella patens consists of a 618 bp open reading frame with high homology (around 72%) to known cox3 sequences of higher plants. Nevertheless, it is a quarter shorter than these. The cox3 gene of P. patens contains no introns and reveals a G+C-content of 41.3%. The region containing the cox3 gene exists as a single copy in the mitochondrial genome as shown by restriction mapping. In the 5′ flanking sequence a putative ribosome binding site and a putative secondary structure were found. Two main transcripts of 2.4 kb and 2.6 kb were detected indicating a complex mitochondrial transcription pattern possibly due to co-transcription. Additional open reading frames were found downstream from, as well as upstream of, the cox3 gene. In Western blots a polyclonal cox3 antibody from yeast detected one single band with an apparent molecular weight of 22 kDa.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: Cytokinin ; Physcomitrella patents (Hedw.) B. S. G. ; Plastid DNA ; Zinc-finger protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plastid DNA of the moss Physcomitrella patens has been sequenced. An open reading frame (ORF 315) was identified downstream from rbcL, between trnR-CCG and psal. This ORF shares homology with zfpA, a putative regulatory gene in Pisum sativum. The moss ORF is preceded by a Shine-Dalgarno sequence, two plastid promoter consensus sequences, and three TATA boxes. A specific probe detected three transcripts of low abundance in the wild-type moss and a cytokininsensitive chloroplast mutant. Steady state levels of zfpA transcripts were different in the two genotypes. In mutant protonemata treated with cytokinin, steady state levels of the largest transcript decreased significantly.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2048
    Keywords: Key words: ATPase ; Mutant (moss) ; Oxygen-evolving complex ; Phosphoglycerate kinase ; Physcomitrella (mutant) ; Plastid division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Cytokinins induce two specific morphological alterations in mosses: (i) the differentiation of a tip-growing cell into a three-faced apical cell (the so-called bud), and (ii) the division of chloroplasts. In a developmental mutant of the moss Physcomitrella patens (Hedw.) B.S.G. (mutant PC22) impeded in both cellular differentiation (bud production) and chloroplast division, addition of cytokinin (N6-Δ2-isopentenyladenine) led to bud production after 3 d in the wild type and after 7 d in the mutant. Hormone induced a division of the mutant macrochloroplasts starting within 24 h and ongoing for 72 h. During this period the abundances of several plastid proteins changed in both genotypes as judged by two-dimensional-protein gel electrophoresis, silver staining and subsequent quantification with novel computer software. Eight of these polypeptides were isolated independently, subjected to microsequencing and thus identified, resulting in the first protein sequence data from a moss. Three polypeptides (24 kDa, 22 kDa, 20 kDa) were found to be homologous to enhancer protein OEE2 of the oxygen-evolving complex, four to represent isoforms of phosphoglycerate kinase (EC 2.7.2.3), and one was identified as the β-chain of chloroplast ATPase (EC 3.6.1.34). Possible involvement of these key enzymes of the chloroplast energy-conversion machinery in organelle division and in cellular differentiation is discussed. Further sequence information was obtained from both subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39). Amounts of these polypeptides were not appreciably affected by cytokinin in moss chloroplasts.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 244 (1994), S. 352-359 
    ISSN: 1617-4623
    Keywords: Archegoniate ; Differentiation mutant ; Plant nuclear DNA ; Repetitive DNA ; Somatic hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A wild-type (WT) strain of the moss Physcomitrella patens (Hedw.) B.S.G., two mutants derived from it (PC22 and P24), and a somatic hybrid, PC22(+)P24, were analysed. Staining of metaphases revealed 54±2 chromosomes in the somatic hybrid and 27 chromosomes in the wild type and the two mutants. Using flow cytometry (FCM), DNA contents were calculated to be 0.6 pg (WT, PC22), 1.2 pg (P24), and 1.6 pg (PC22(+)P24) per nucleus, respectively. Southern hybridization provided evidence for at least one family of highly repetitive DNA and, furthermore, revealed different amounts of repetitive DNA in the four genotypes. However, these sequences cannot account for the 100% increase in the nuclear DNA amount in mutant P24, relative to wild type. In FCM analyses every moss geno-type generated just one single peak of fluorescence, indicating an arrest in the cell cycle during the daytime. Thermal denaturation of wild-type DNA revealed a G+C content of 34.6% for total DNA and 38.6% for plastid DNA. A cDNA library of 1.2 × 106 independent clones was established, from which sequences homologous to cab and rbcS, respectively, were isolated. These genes show significant homologies to those of higher plants, and, likewise, comprise multigene families. No restriction fragment length polymorphisms could be detected between the four moss genotypes using these cDNA probes.
    Type of Medium: Electronic Resource
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