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New Insight in Ethylene Signaling： Autokinase Activity of ETR1 Modulates the Interaction of Receptors and EIN2

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Molecular Plant 2010年第5期3卷 882-889页

作者： Melanie M.A. Bisson Georg Groth Heinrich-Heine Universitat Dusseldorf Biochemie der Pflanzen Universitatsstr. 1 40225 Dosseldorf Germany

Ethylene insensitive 2 （EIN2）, an integral membrane protein of the ER network, has been identified as the central regulator of the ethylene signaling pathway. Still, the mechanism by which the ethylene signal is transferred from the receptors to EIN2 has not been solved yet. Here, we show that protein phosphorylation is a key mechanism to control the interaction of EIN2 and the receptors. In vivo and in vitro fluorescence studies reveal that the kinase domain of the receptors is essential for the interaction. Cyanide, an ethylene agonist, which is known to reduce auto-phosphorylation of the ethylene receptor ethylene resistant 1 （ETR1） or a mutation in the kinase domain of ETR1 that prevents autophosphorylation （H353A）, increases the affinity of the receptors for EIN2. On the other hand, mimicking permanent auto-phosphorylation of ETR1 as in the mutant H353E releases the EiN2-ETR1 interaction from the control by the plant hormone. Based on our data, we propose a novel model on the integration of EIN2 in the ethylene signaling cascade.

关键词： Membrane biochemistry protein phosphorylation/dephosphorylation receptors signal transduction membrane proteins ethylene protein-protein interaction.

Pitt, a Novel Tetratricopeptide Repeat Protein Involved in Light-Dependent Chlorophyll Biosynthesis and Thylakoid Membrane Biogenesis in Synechocystis sp. PCC 6803

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Molecular Plant 2009年第6期2卷 1289-1297页

作者： Marco Schottkowski Janina Ratke Ulrike Oster Marc Nowaczyk Jorg Nickelsen Molekulare pflanzenwissenschaften Biozentrum LMU Munchen Groβhaderner Str. 2-4 82152 Planegg-Martinsried Germany biochemie und Physiologie der pflanzen Biozentrum LMU Munchen Groβhaderner Str. 2-4 82152 Planegg-Martinsried Germany ls biochemie der pflanzen Ruhr-Universitait Bochum Universitatsstr. 150 44780 Bochum Germany

Biogenesis of photosynthetic pigment/protein complexes is a highly regulated process that requires various assisting factors. Here, we report on the molecular analysis of the Pitt gene （sir1644） from the cyanobacterium Synechocystis sp. PCC 6803 （Synechocystis 6803） that encodes a membrane-bound tetratricopeptide repeat （TPR） protein of formerly unknown function. Targeted inactivation of Pitt affected photosynthetic performance and light-dependent chlorophyll synthesis. Yeast two-hybrid analyses and native PAGE strongly suggest a complex formation between Pitt and the light-dependent protochlorophyllide oxidoreductase （POR）. Consistently, POR levels are approximately threefold reduced in the pitt insertion mutant. The membrane sublocalization of Pitt was found to be dependent on the presence of the periplasmic photosystem Ⅱ （PSⅡ） biogenesis factor PratA, supporting the idea that Pitt is involved in the early steps of photosynthetic pigment/protein complex formation.

关键词： Synechocystis TPR protein POR thylakoid membrane chlorophyll.

Proteomic Analysis of the Proplastid Envelope Membrane Provides Novel Insights into Small Molecule and Protein Transport across Proplastid Membranes

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Molecular Plant 2009年第6期2卷 1247-1261页

作者： Andrea Brautigam Andreas RM. Weber Institut fur biochemie der pflanzen Heinrich Heine Universitat Universitatsstrasse 1 D-40225 Dusseldorf Germany Genetics Program Michigan State University East Lansing MI USA Department of Plant Biology Michigan State University East Lansing MI USA

Proplastids are undifferentiated plastids of meristematic tissues that synthesize amino acids for protein synthesis, fatty acids for membrane lipid production, and purines and pyrimidines for DNA and RNA synthesis. Unlike chloroplasts, proplastids depend on supply, with reducing power, energy, and precursor metabolites from the remainder of the cell. Comparing proplastid and chloroplast envelope proteomes and the corresponding transcriptomes of leaves and shoot apex revealed a clearly distinct composition of the proplastid envelope. It is geared towards import of metabolic precursors and export of product metabolites for the rapidly dividing cell. The analysis also suggested a new role for the triosephosphate translocator in meristematic tissues, identified the route of organic nitrogen import into proplastids, and detected an adenine nucleotide exporter. The protein import complex contains the import receptors Toc120 and Toc132 and lacks the redox sensing complex subunits of Tic32, Tic55, and Tic62, which mirrors the expression patterns of the corresponding genes in leaves and the shoot apex. We further show that the protein composition of the internal membrane system is similar to etioplasts, as it is dominated by the ATP synthase complex and thus remarkably differs from that of chloroplast thylakoids.

关键词： Membrane biochemistry proteomics transporters chloroplast biology membrane proteins transcriptome analysis.

Proteomic and molecular investigations revealed that Acidithiobacillus caldus adopts multiple strategies for adaptation to NaCl stress

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Chinese Science Bulletin 2014年第3期59卷 301-309页

作者： Xu Guo Chengying Jiang Yuanming Luo Mingjiang Zhang Ansgar Poetsch Shuangjiang Liu State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of Sciences University of Chinese Academy of Sciences Environmental Microbiology and Biotechnology Research Center Institute of MicrobiologyChinese Academy of Sciences Lehrstuhl for biochemie der pflanzen Ruhr Universitaet

Acidithiobacillus caldus plays an important role in commercial *** understand how NaCl stress adaptation occurs in ***,we grew *** strain SM-1 in media containing high NaCl ***-1 grew at concentrations of up to 1.0-mol L^(-1)NaCl,but growth was severely inhibited at higher *** analysis showed that SM-1 used multiple strategies to respond to NaCl *** addition to several heatshock proteins,enzymes involved in proline biosynthesis increased under NaCl *** addition,two DNA-binding proteins and a third protein of unknown function(Atc_(1291)),which was subsequently identified as a putative single-stranded DNA-binding protein,were up-regulated in the presence of NaCl *** DNA-binding proteins might play a role in response to osmotic ***_(1291)was cloned and expressed in Escherichia ***,we found that *** BL21/pET28a-atc_(1291)grew to higher cell densities than *** BL21/pET28a,regardless of NaCI *** to Atc_(1291)were identified in several groups of *** role of Atc_(1291)in enhancing cell growth needs further investigation.

关键词：蛋白质组学盐胁迫 DNA结合蛋白 NaCl胁迫显示分子抑制浓度大肠杆菌

The Plastid Envelope CHLOROPLAST MANGANESE TRANSPORTER1 Is Essential for Manganese Homeostasis in Arabidopsis

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Molecular Plant 2018年第7期11卷 955-969页

作者： Marion Eisenhut Natalie Hoecker Sidsel Birkelund Schmidt Rubek Merina Basgaran Samantha Flachbart Peter Jahns Tabea Eser Stefan Geimer Seren Husted Andreas P.M. Weber Dario Leister Anja Schneider biochemie der pflanzen Cluster of Excellence on Plant Science (CEPLAS) Heinrich-Heine-University Di3sseldorf 40225 Desseldorf Germany Molekularbiologie der pflanzen (Botanik) Department Biologie I Ludwig-Maximilians-University Mernchen 82152 Martinsried Germany Department of Plant and Environmental Sciences and Copenhagen Plant Science Centre (CPSC) Faculty of Science University of Copenhagen 1871 Frederiksberg Denmark Zellbiologie/Elektronenmikroskopie NW I/B1 Universit&t Bayreuth 95447 Bayreuth Germany These authors contributed equally to this article.

The transition metal manganese （Mn） is indispensable for photoautotrophic growth since photosystem II （PSII） employs an inorganic Mn4CaOs cluster for water splitting. Here, we show that the Arabidopsis membrane protein CHLOROPLAST MANGANESE TRANSPORTER1 （CMT1） is involved in chloroplast Mn homeostasis. CMT1 is the closest homolog of the previously characterized thylakoid Mn transporter PHOTOSYNTHESIS-AFFECTED MUTANT71 （PAM71）. In contrast to PAM71, CMT1 resides at the chloro- plast envelope and is ubiquitously expressed. Nonetheless, like PAM71, the expression of CMT1 can also alleviate the Mn-sensitive phenotype of yeast mutant qomrl. The cmtl mutant is severely suppressed in growth, chloroplast ultrastructure, and PSII activity owing to a decrease in the amounts of pigments and thylakoid membrane proteins. The importance of CMT1 for chloroplast Mn homeostasis is demonstratedby the significant reduction in chloroplast Mn concentrations in cmtl-1, which exhibited reduced Mn binding in PSII complexes. Moreover, CMT1 expression is downregulated in Mn-surplus conditions. The pam71 cmtl-ldouble mutant resembles the cmtl-f single mutant rather than pare71 in most respects. Taken together, our results suggest that CMT1 mediates Mn2 uptake into the chloroplast stroma, and that CMT1 and PAM71 function sequentially in Mn delivery to PSII across the chloroplast envelope and the thylakoid membrane.

关键词： manganese transporter envelope chloroplast photosynthesis Arabidopsis

NITROGEN FIXATION OF Azorhizobium IN ARTIFICIALLY INDUCED ROOT PARA-NODULES IN WHEAT

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Science China Chemistry 1992年第12期35卷 1463-1470页

作者：陈廷伟 SIEGFRIED SCHERER PETER BGER Soils and Fertilizer Institute Chinese Academy of Agriculiural SciencesBeijing 100081PRC Bacteriological Institute Faculty of Agriculture Technical University Munchen D-8050 Freising Germany Lehrstuhl für Physiologie und biochemie der pflanzen Universitat Konstanz D-7750 Konstanz Germany

Nodule-like structures (para-nodules) can be induced in wheat roots by low concentrationsof 2,4-dichlorophenoxyacetic acid (2,4-D), a synthetic analogue of the plant growth hor-mone indole-acetic acid. Growth of nodulated wheat plants was not affected by 2,4-D treat-ment. Infection of these?para-nodules by adding oxygen-tolerant Azorhizobium caulinodans isolat-ed from stem nodules of the tropical legume Sesbania rostrata resulted in massive prolifera-tion of bacterial cells in the intercellular spaces as well as inside the para-nodular ***-nodules colonized by Azorhizobium coulinodans not only reduced acetylene, but alsotransferred fixed nitrogen to the plant, contributing up to 16--23% of the nitrogen budgetof infected wheat.

关键词： Azorhizobium nitrogen fixation nodules wheat

The Arabidopsis Protein CGLD11 Is Required for Chloroplast ATP Synthase Accumulation

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Molecular Plant 2016年第6期9卷 885-899页

作者： Sabine Grahl Bennet Reiter Irene Luise Gugel Evgenia Vamvaka Chiara Gandini Peter Jahns Jurgen Soil Dario Leister Thilo Ruhle Lehrstuhl fur biochemie und Physiologie der pflanzen (Botanik) Department Biologie I Ludwig-Maximilians-Universitat Munchen 82152 Planegg-MartinsriedGermany Lehrstuhl for Molekularbiologie der pflanzen (Botanik) Department Biologie I Ludwig-MaximHians-Universitat MQnchen 82152 Planegg-Martinsried Germany Plant Biochemistry Heinrich-Heine-University DLisseldorf Universittsstrasse 1 40225 Desseldorf Germany Munich Centre for Integrated Protein Science CiPSM Ludwig-Maximilians Universitat Menchen Butenandtstr. 5 - 13 81377 Munich Germany These authors contributed equally to this article.

ATP synthases in chloroplasts （cpATPase） and mitochondria （mtATPase） are responsible for ATP production during photosynthesis and oxidative phosphorylation, respectively. Both enzymes consist of two multi- subunit complexes, the membrane-bound coupling factor O and the soluble coupling factor 1. During cpATPase biosynthesis, several accessory factors facilitate subunit production and orchestrate complex assembly. Here, we describe a new auxiliary protein in Arabidopsis thaliana, which is required for cpATPase accumulation. AtCGLD11 （CONSERVED IN THE GREEN LINEAGE AND DIATOMS 11） is a protein without any known functional domain and shows dual localization to chloroplasts and mitochondria. Loss of AtCGLDll function results in reduced levels of cpATPase and impaired photosynthetic performance with lower rates of ATP synthesis. In yeast two-hybrid experiments, AtCGLD11 interacts with the 13 subunits of the cpATPase and mtATPase. Our results suggest that AtCGLD11 functions in F1 assembly during cpATPase biogenesis, while its role in mtATPase biosynthesis may not, or not yet, be essential.

关键词： chloroplast mitochondria photosynthesis dual targeting ATP synthase assembly

Heat Shock Factors HsfB 1 and HsfB2b Are Involved in the Regulation of Pdf1.2 Expression and Pathogen Resistance in Arabidopsis

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Molecular Plant 2009年第1期2卷 152-165页

作者： Mukesh Kumar Wolfgang Busch Hannah Birke Birgit Kemmerling Thorsten Nurnberger Friedrich Schoffl Present address: Heinrich-Pette-lnstitut Martinistrabe 52 D-20251 Hamburg Germany Max-Planck-lnstitut fur Entwicklungsbiologie Abteilung Molekularbiologie Speemannstrabe 37-39 D-72076 Tubingen Germany Universitat Tubingen Zentrum for Molekularbiologie der Pflanzen (ZMBP)--AIIgemeine Genetik Auf der Morgenstelle 28 D-72076 Tubingen Germany Universitat Tubingen Zentrum fur Molekularbiologie der Pflanzen (ZMBP)--Biochemie der Pflanzen Auf der Morgenstelle 5 D-72076 Tubingen Germany

In order to assess the functional roles of heat stress-induced class B-heat shock factors in Arabidopsis, we investigated T-DNA knockout mutants of AtHsfB1 and AtHsfB2b. Micorarray analysis of double knockout hsfB1/hsfB2b plants revealed as strong an up-regulation of the basal mRNA-levels of the defensin genes Pdfl.2a/b in mutant plants. The Pdfexpression was further enhanced by jasmonic acid treatment or infection with the necrotrophic fungus Alternaria brassicicola. The single mutant hsfB2b and the double mutant hsfB 1/B2b were significantly improved in disease resistance after A. brassicicola infection. There was no indication for a direct interaction of Hsf with the promoter of Pdf1.2, which is devoid of perfect HSE consensus Hsf-binding sequences. However, changes in the formation of late HsfA2-dependent HSE binding were detected in hsfB1/B2b plants. This suggests that HsfB1/B2b may interact with class A-Hsf in regulating the shut-off of the heat shock response. The identification of Pdfgenes as targets of Hsf-dependent negative regulation is the first evidence for an interconnection of Hsf in the regulation of biotic and abiotic responses.

关键词： Abiotic/environmental stress transcriptional control and transcription factors transcriptome analysis defense responses disease resistance Arabidopsis.

Transgenic Poplar Plants for the Investigation of ABA-Dependent Salt and Drought Stress Adaptation in Trees

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American Journal of Plant Sciences 2016年第9期7卷 1337-1356页

作者： Domenica Hamisch David Kaufholdt Jennifer-Christin Kuchernig Florian Bittner Ralf R. Mendel Robert Hänsch Jennifer Popko Institut für pflanzenbiologie Technische Universitat Braunschweig Braunschweig Germany Albrecht-von-Haller-Institutfür pflanzenwissenschaften Abteilung Biochemie der Pflanzen Georg-August-Universitat Gottingen Germany

Important functions of the plant hormone abscisic acid (ABA) in stress reactions, growth and photosynthetic processes are extensively studied in the model plant Arabidopsis thaliana. This paper investigates the importance of Moco-sulphurase ABA3 and aldehyde oxidase (AO) on ABA-biosynthesis in Populus × canescens. ABA3 is essential for activation of the molybdenum enzymes AO and xanthine dehydrogenase (XDH). AO itself catalyzes the last step in ABA-biosynthesis. Generation of transgenic poplar plants altered in ABA3 and AO-activity using RNAi knock down and overexpression was performed. Whereas RNAi-AO plants show a specific loss of AO activity, the RNAi-ABA3 plants has a strongly reduced activity of both molybdenum enzymes: AO and XDH. Constructs of AO and ABA3-promoters fused to β-glucuronidase provide the basis to investigate transcriptional regulation of ABA-biosynthetic processes under stress conditions. Application of high salt concentrations and different drought stress intensities does change the endogenous AO or XDH neither on the side of transcription nor on protein activity. On phytohormone level however, water loss leads to increased ABA-amounts regardless of whether transgenic or wildtype plants are studied. Salt application resulted in higher ABA-levels in all analyzed plant lines. The down regulation of AO in the two different RNAi-plant lines strongly prevented a wildtype-like increase of ABA-levels. Whereas the WT plants accumulated up to 6000 ng ABA g-1 FW-1 after 16 h of salt stress exposure, plants of the RNAi lines revealed a markedly lower increase of only up to 2000 ng ABA g-1 FW-1. Opposing to these observations, ABA-levels increased during drought without any influence by the RNAi-effect. These results revealed that although stresses did not result in a visible increased AO-activity, ABA-production was influenced by AO and ABA3 at least under salinity.

关键词： ABA Biosynthesis Drought Stress Populus × canescens Transgenic Poplar Salt Stress