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Publikationen - Molekulare Signalverarbeitung

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Publikation

Floková, K.; Feussner, K.; Herrfurth, C.; Miersch, O.; Mik, V.; Tarkowská, D.; Strnad, M.; Feussner, I.; Wasternack, C.; Novák, O.; A previously undescribed jasmonate compound in flowering Arabidopsis thaliana – The identification of cis-(+)-OPDA-Ile Phytochemistry 122, 230-237, (2016) DOI: 10.1016/j.phytochem.2015.11.012

Jasmonates (JAs) are plant hormones that integrate external stress stimuli with physiological responses. (+)-7-iso-JA-L-Ile is the natural JA ligand of COI1, a component of a known JA receptor. The upstream JA biosynthetic precursor cis-(+)-12-oxo-phytodienoic acid (cis-(+)-OPDA) has been reported to act independently of COI1 as an essential signal in several stress-induced and developmental processes. Wound-induced increases in the endogenous levels of JA/JA-Ile are accompanied by two to tenfold increases in the concentration of OPDA, but its means of perception and metabolism are unknown. To screen for putative OPDA metabolites, vegetative tissues of flowering Arabidopsis thaliana were extracted with 25% aqueous methanol (v/v), purified by single-step reversed-phase polymer-based solid-phase extraction, and analyzed by high throughput mass spectrometry. This enabled the detection and quantitation of a low abundant OPDA analog of the biologically active (+)-7-iso-JA-L-Ile in plant tissue samples. Levels of the newly identified compound and the related phytohormones JA, JA-Ile and cis-(+)-OPDA were monitored in wounded leaves of flowering Arabidopsis lines (Col-0 and Ws) and compared to the levels observed in Arabidopsis mutants deficient in the biosynthesis of JA (dde2-2, opr3) and JA-Ile (jar1). The observed cis-(+)-OPDA-Ile levels varied widely, raising questions concerning its role in Arabidopsis stress responses.
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

Wasternack, C.; Miersch, O.; Kramell, R.; Hause, B.; Ward, J.; Beale, M.; Boland, W.; Parthier, B.; Feussner, I.; Jasmonic acid: biosynthesis, signal transduction, gene expression Fett/Lipid 100, 139-146, (1998) DOI: 10.1002/(SICI)1521-4133(19985)100:4/5<139::AID-LIPI139>3.0.CO;2-5

Jasmonic acid (JA) is an ubiquitously occurring plant growth regulator which functions as a signal of developmentally or environmentally regulated expression of various genes thereby contributing to the defense status of plants [1–5]. The formation of jasmonates in a lipid‐based signalling pathway via octadecanoids seems to be a common principle for many plant species to express wound‐ and stressinduced genes [4, 5].There are various octadecanoid‐derived signals [3]. Among them, jasmonic acid and its amino acid conjugates are most active in barley, supporting arguments that β‐oxidation is an essential step in lipid‐based JA mediated responses. Furthermore, among derivatives of 12‐oxophytodienoic acid (PDA) carrying varying length of the carboxylic acid side‐chain, only those with a straight number of carbon atoms are able to induce JA responsive genes in barley leaves after treatment with these compounds. Barley leaves stressed by treatment with sorbitol solutions exhibit mainly an endogenous rise of JA and JA amino acid conjugates suggesting that both of them are stress signals. Data on organ‐ and tissue‐specific JA‐responsive gene expression will be presented and discussed in terms of “JA as a master switch” among various lipid‐derived signals.
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

Hause, B.; Feussner, K.; Wasternack, C.; Nuclear Location of a Diadenosine 5′,5′”-P1,P4Tetraphosphate (Ap4A) Hydrolase in Tomato Cells Grown in Suspension Cultures Bot. Acta 110, 452-457, (1997) DOI: 10.1111/j.1438-8677.1997.tb00662.x

Diadenosine 5′,5′”‐P1,P4‐tetraphosphate (Ap4A) cleaving enzymes are assumed to regulate intracellular levels of Ap4A, a compound known to affect cell proliferation and stress responses. From plants an Ap4A hydrolase was recently purified using tomato cells grown in suspension. It was partially sequenced and a peptide antibody was prepared (Feussner et al., 1996). Using this polyclonal monospecific antibody, an abundant nuclear location of Ap4A hydrolase in 4‐day‐old cells of atomato cell suspension culture is demonstrated here by means of immunocytochemical techniques using FITC (fluorescein‐5‐isothiocyanate) labeled secondary antibodies. The microscopic analysis of the occurrence of Ap4A hydrolase performed for different stages of the cell cycle visualized by parallel DAPI (4,6‐diamidino‐2‐phenylindole) staining revealed that the protein accumulates within nuclei of cells in the interphase, but is absent in the nucleus as well as cytoplasm during all stages of mitosis. This first intracellular localization of an Ap4A degrading enzyme within the nucleus and its pattern of appearance during the cell cycle is discussed in relation to the suggested role of Ap4A in triggering DNA synthesis and cell proliferation.
Publikation

Feussner, K.; Feussner, I.; Leopold, I.; Wasternack, C.; Isolation of a cDNA coding for an ubiquitin-conjugating enzyme UBC1 of tomato - the first stress-induced UBC of higher plants FEBS Lett. 409, 211-215, (1997) DOI: 10.1016/S0014-5793(97)00509-7

A clone of an ubiquitin‐conjugating enzyme (UBC) was isolated from a λ‐ZAP‐cDNA library, generated from mRNA of tomato (Lycopersicon esculentum) cells grown in suspension for 3 days. The open reading frame called Le UBC1, encodes for a polypeptide with a predicted molecular mass of 21.37 kDa, which was confirmed by bacterial overexpression and SDS‐PAGE. Database searches with Le UBC1 showed highest sequence similarities to UBC1 of bovine and yeast. By Southern blot analysis Le UBC1 was identified as a member of a small E2 subfamily of tomato, presumably consisting of at least two members. As revealed by Northern blot analysis Le UBC1 is constitutively expressed in an exponentially growing tomato cell culture. In response to heat shock an increase in Le UBC1‐mRNA was detectable. A strong accumulation of the Le UBC1‐transcript was observed by exposure to heavy metal stress which was performed by treatment with cadmium chloride (CdCl2). The cellular uptake of cadmium was controlled via ICP‐MS measurements. The data suggest that like in yeast, in plants a certain subfamily of UBC is specifically involved in the proteolytic degradation of abnormal proteins as result of stress.
Publikation

Feussner, K.; Guranowski, A.; Kostka, S.; Wasternack, C.; Diadenosine 5′,5‴- P1,P4-tetraphosphate (Ap4A) Hydrolase from Tomato (Lycopersicon esculentum cv. Lukullus) -Purification, Biochemical Properties and Behaviour during Stress Z. Naturforsch. C 51, 477-486, (1996) DOI: 10.1515/znc-1996-7-805

Dinucleoside 5′,5‴-P1,P4-tetraphosphate hydrolase (EC 3.6.1.17) has been purified to homogeneity from tomato (Lycopersicon esculentum) cells grown in suspension. The purification procedure comprised ammonium sulphate fractionation following five standard chroma­ tography steps and a final chromatography on Ap4A-Sepharose.
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