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Publikationen - Molekulare Signalverarbeitung

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Publikation

Otto, M.; Naumann, C.; Brandt, W.; Wasternack, C.; Hause, B.; Activity Regulation by Heteromerization of Arabidopsis Allene Oxide Cyclase Family Members Plants 5, 3, (2016) DOI: 10.3390/plants5010003

Jasmonates (JAs) are lipid-derived signals in plant stress responses and development. A crucial step in JA biosynthesis is catalyzed by allene oxide cyclase (AOC). Four genes encoding functional AOCs (AOC1, AOC2, AOC3 and AOC4) have been characterized for Arabidopsis thaliana in terms of organ- and tissue-specific expression, mutant phenotypes, promoter activities and initial in vivo protein interaction studies suggesting functional redundancy and diversification, including first hints at enzyme activity control by protein-protein interaction. Here, these analyses were extended by detailed analysis of recombinant proteins produced in Escherichia coli. Treatment of purified AOC2 with SDS at different temperatures, chemical cross-linking experiments and protein structure analysis by molecular modelling approaches were performed. Several salt bridges between monomers and a hydrophobic core within the AOC2 trimer were identified and functionally proven by site-directed mutagenesis. The data obtained showed that AOC2 acts as a trimer. Finally, AOC activity was determined in heteromers formed by pairwise combinations of the four AOC isoforms. The highest activities were found for heteromers containing AOC4 + AOC1 and AOC4 + AOC2, respectively. All data are in line with an enzyme activity control of all four AOCs by heteromerization, thereby supporting a putative fine-tuning in JA formation by various regulatory principles.
Publikation

Wasternack, C.; Stenzel, I.; Hause, B.; Hause, G.; Kutter, C.; Maucher, H.; Neumerkel, J.; Feussner, I.; Miersch, O.; The wound response in tomato – Role of jasmonic acid J. Plant Physiol. 163, 297-306, (2006) DOI: 10.1016/j.jplph.2005.10.014

Plants respond to mechanical wounding or herbivore attack with a complex scenario of sequential, antagonistic or synergistic action of different signals leading to defense gene expression. Tomato plants were used as a model system since the peptide systemin and the lipid-derived jasmonic acid (JA) were recognized as essential signals in wound-induced gene expression. In this review recent data are discussed with emphasis on wound-signaling in tomato. The following aspects are covered: (i) systemin signaling, (ii) JA biosynthesis and action, (iii) orchestration of various signals such as JA, H2O2, NO, and salicylate, (iv) local and systemic response, and (v) amplification in wound signaling. The common occurrence of JA biosynthesis and systemin generation in the vascular bundles suggest JA as the systemic signal. Grafting experiments with JA-deficient, JA-insensitive and systemin-insensitive mutants strongly support this assumption.
Publikation

Bücking, H.; Förster, H.; Stenzel, I.; Miersch, O.; Hause, B.; Applied jasmonates accumulate extracellularly in tomato, but intracellularly in barley FEBS Lett. 562, 45-50, (2004) DOI: 10.1016/S0014-5793(04)00178-4

Jasmonic acid (JA) and its derivatives are well‐characterized signaling molecules in plant defense and development, but the site of their localization within plant tissue is entirely unknown. To address the question whether applied JA accumulates extracellularly or intracellularly, leaves of tomato and barley were fed with 14C‐labeled JA and the label was localized in cryofixed and lyophilized leaf tissues by microautoradiography. In tomato the radioactivity was detectable within the apoplast, but no label was found within the mesophyll cells. By contrast, in barley leaf tissues, radioactivity was detected within the mesophyll cells suggesting a cellular uptake of exogenously applied JA. JA, applied to leaves of both plants as in the labeling experiments, led in all leaf cells to the expression of JA‐inducible genes indicating that the perception is completed by JA signal transduction.
Publikation

Nibbe, M.; Hilpert, B.; Wasternack, C.; Miersch, O.; Apel, K.; Cell death and salicylate- and jasmonate-dependent stress responses in Arabidopsis are controlled by single cet genes Planta 216, 120-128, (2002) DOI: 10.1007/s00425-002-0907-1

The jasmonic acid (JA)-dependent regulation of the Thi2.1 gene had previously been exploited for setting up a genetic screen for the isolation of signal transduction mutants of Arabidopsis thaliana (L.) Heynh. that constitutively express the thionin gene. Several cet mutants had been isolated which showed a constitutive expression of the thionin gene. These cet mutants, except for one, also showed spontaneous leaf cell necrosis and were up-regulated in the expression of the PR1 gene, reactions often associated with the systemic acquired resistance (SAR) pathway. Four of these cet mutants, cet1, cet2, cet3 and cet4.1 were crossed with the fad triple and coi1 mutants that are blocked at two steps within the JA-dependent signaling pathway, and with transgenic NahG plants that are deficient in salicylic acid (SA) and are unable to activate SAR. Analysis of the various double-mutant lines revealed that the four cet genes act within a signaling cascade at or prior to branch points from which not only JA-dependent signals but also SA-dependent signaling and cell death pathways diverge.
Publikation

Hause, B.; Vörös, K.; Kogel, K.-H.; Besser, K.; Wasternack, C.; A Jasmonate-responsive Lipoxygenase of Barley Leaves is Induced by Plant Activators but not by Pathogens J. Plant Physiol. 154, 459-462, (1999) DOI: 10.1016/S0176-1617(99)80283-1

Using the recently isolated eDNA clone LOX2 : Hv : 1 which codes for the most abundant jasmonateinducible lipoxygenase (LOX) in barley leaves (Vörös et al., 1998), we analysed the capability of different activators of systemic activated resistance (SAR) to induce the expression of that LOX. Upon treatment of barley leaves with salicylate, 2,6-dichloroisonicotinic acid and benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester, all these compounds were able to induce the expression of the LOX2 : Hv : 1 gene, whereas upon infection with the powdery mildew fungus (Blumeria graminis f. sp. hordei) mRNA accumulation was not detectable in compatible or in incompatible interactions. The induction of the LOX2 : Hv : 1 protein by SAR activators and the expression of different sets of genes induced by jasmonate and salicylate, respectively, are discussed in relation to defense responses against pathogenic fungi.
Publikation

Bohlmann, H.; Vignutelli, A.; Hilpert, B.; Miersch, O.; Wasternack, C.; Apel, K.; Wounding and chemicals induce expression of the Arabidopsis thaliana gene Thi2.1, encoding a fungal defense thionin, via the octadecanoid pathway FEBS Lett. 437, 281-286, (1998) DOI: 10.1016/S0014-5793(98)01251-4

In seedlings of Arabidopsis thaliana the thionin gene Thi2.1 is inducible by methyl jasmonate, wounding, silver nitrate, coronatine, and sorbitol. We have used a biochemical and genetic approach to test the signal transduction of these different inducers. Both exogenously applied jasmonates and jasmonates produced endogenously upon stress induction, lead to GUS expression in a Thi2.1 promoter-uidA transgenic line. No GUS expression was observed in a coi1 mutant background which lacks jasmonate perception whereas methyl jasmonate and coronatine but not the other inducers were able to overcome the block in jasmonic acid production in a fad3-2 fad7-2 fad8 mutant background. Our results show conclusively that all these inducers regulate Thi2-1 gene expression via the octadecanoid pathway.
Publikation

Wasternack, C.; Ortel, B.; Miersch, O.; Kramell, R.; Beale, M.; Greulich, F.; Feussner, I.; Hause, B.; Krumm, T.; Boland, W.; Parthier, B.; Diversity in octadecanoid-induced gene expression of tomato J. Plant Physiol. 152, 345-352, (1998) DOI: 10.1016/S0176-1617(98)80149-1

In tomato plants wounding leads to up-regulation of various plant defense genes via jasmonates (Ryan, 1992; Bergey et al., 1996). Using this model system of jasmonic acid (JA) signalling, we analyzed activity of octadecanoids to express JA-responsive genes. Leaf treatments were performed with naturally occurring octadecanoids and their molecular mimics such as coronatine or indanone conjugates. JA responses were recorded in terms of up- or down-regulation of various genes by analyzing transcript accumulation, and at least partially in vitro translation products and polypeptide pattern of leaf extracts. The data suggest: (i) 12-Oxo-phytodienoic acid and other intermediates of the octadecanoid pathway has to be ß-oxidized to give a JA response, (ii) Octadecanoids which can not be ß-oxidized are inactive, (iii) JA, its methyl ester (JM), and its amino acid conjugates are most active signals in tomato leaves leading to up regulation of mainly wound-inducible genes and down-regulation of mainly <house-keeping> genes, (iv) Some compounds carrying a JA/JM- or JA amino acid conjugate-like structure induce/repress only a subset of genes suggesting diversity of JA signalling.
Publikation

Ratajczak, R.; Feussner, I.; Hause, B.; Böhm, A.; Parthier, B.; Wasternack, C.; Alteration of V-type H+-ATPase during methyljasmonate-induced senescence in barley (Hordeum vulgare L. cv. Salome) J. Plant Physiol. 152, 199-206, (1998) DOI: 10.1016/S0176-1617(98)80133-8

In barley leaves, the application of (−)-jasmonic acid or its methyl ester (JAME) induces a senescencelike phenotype. This is accompanied by the synthesis of abundant proteins, so-called jasmonate-induced proteins (JlPs). Here, we show that modifications of vacuolar H+-ATPase (V-ATPase) subunits are jasmo-nate inducible. Using immunofluorescence analysis, we demonstrate that V-ATPase of barley leaves is exclusively located at the tonoplast also upon JAME treatment. Total ATP-hydrolysis activity of microsomal fractions increased by a factor of 10 during 72 h of JAME-treatment, while Bafilomycin Ai-sensitive ATP-hydrolysis activity, which is usually referred to V-ATPase activity, increased by a factor of about 2 in tono-plast-enriched membrane fractions. Moreover, due to JAME treatment there was a pronounced increase in ATP-hydrolysis activity at pH 6.2. This activity was not affected by inhibitors of P-, F-, or V-ATPases. However, biochemical analysis of partially purified V-ATPase suggests, that this activity might be due at least in part to the V-ATPase. JAME-treatment seems to change biochemical properties of the V-ATPase, i.e. a shift of the pH optimum of activity to a more acidic pH and a decrease in Bafilomycin A1 sensitivity. This is accompanied by the appearance of several additional forms of V-ATPase subunits which might represent either different isoforms or post-translationally modified proteins. We suggest that these changes in properties of the V-ATPase, which is involved in house-keeping and stress responses, may be due to JAME-induced senescence to overcome concomitant changes of the vacuolar membrane.
Publikation

Churin, J.; Hause, B.; Feussner, I.; Maucher, H. P.; Feussner, K.; Börner, T.; Wasternack, C.; Cloning and expression of a new cDNA from monocotyledonous plants coding for a diadenosine 5′,5′′′-P1,P4-tetraphosphate hydrolase from barley (Hordeum vulgare) FEBS Lett. 431, 481-485, (1998) DOI: 10.1016/S0014-5793(98)00819-9

From a cDNA library generated from mRNA of white leaf tissues of the ribosome‐deficient mutant ‘albostrians' of barley (Hordeum vulgare cv. Haisa) a cDNA was isolated carrying 54.2% identity to a recently published cDNA which codes for the diadenosine‐5′,5′′′‐P1,P4‐tetraphosphate (Ap4A) hydrolase of Lupinus angustifolius (Maksel et al. (1998) Biochem. J. 329, 313–319), and 69% identity to four partial peptide sequences of Ap4A hydrolase of tomato. Overexpression in Escherichia coli revealed a protein of about 19 kDa, which exhibited Ap4A hydrolase activity and cross‐reactivity with an antibody raised against a purified tomato Ap4A hydrolase (Feussner et al. (1996) Z. Naturforsch. 51c, 477–486). Expression studies showed an mRNA accumulation in all organs of a barley seedling. Possible functions of Ap4A hydrolase in plants will be discussed.
Publikation

Kramell, R.; Miersch, O.; Hause, B.; Ortel, B.; Parthier, B.; Wasternack, C.; Amino acid conjugates of jasmonic acid induce jasmonate-responsive gene expression in barley (Hordeum vulgare L.) leaves FEBS Lett. 414, 197-202, (1997) DOI: 10.1016/S0014-5793(97)01005-3

Leaves of barley (Hordeum vulgare L. cv. Salome ) treated with jasmonic acid (JA), its methyl ester (JM), or its amino acid conjugates exhibit up‐regulation of specific genes and down‐regulation of house‐keeping genes. This transcriptional regulation exhibits several specificities. (i) The (−)‐enantiomers are more active, and conjugates are mainly active if they carry an l ‐amino acid moiety. (ii) The various JA‐responsive genes respond differentially to enantiomeric and chiralic forms. (iii) Both JA and its amino acid conjugates exhibiting no or negligible interconversion induce/repress genes.
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