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Chloroplast Proteins without Cleavable Transit Peptides： Rare Exceptions or a Major Constituent of the Chloroplast Proteome？

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Molecular Plant 2009年第6期2卷 1325-1335页

作者： Ute Armbruster Alexander Hertle Elina Makarenko Jessica Zuhlke Mathias Pribil Angela Dietzmann ivo Schliebner Elena Aseeva Elena Fenino Michael Scharfenberg Christian Voigt Dario Leister Lehrstuhl fur botanik Department Biologie I Ludwig-Maximilians-Universit~t MQnchen Menzinger Str. 67 D-80638 Munchen Germany

Most chloroplast proteins （cp proteins） are nucleus-encoded, synthesized on cytosolic ribosomes as precursor proteins containing a presequence （cTP）, and post-translationally imported via the Tic/Toc complex into the organelle, where the cTP is removed. Only a few unambiguous instances of cp proteins that do not require cTPs （non-canonical cp proteins） have been reported so far. However, the survey of data from large-scale proteomic studies presented here suggests that the fraction of such proteins in the total cp proteome might be as large as -30%. To explore this discrepancy, we chose a representative set of 28 putative non-canonical cp proteins, and used in vitro import and Red Fluorescent Protein （RFP）-fusion assays to determine their sub-cellular destinations. Four proteins, including embryo defective 1211, glycolate oxidase 2, protein disulfide isomerase-like protein （PDii）, and a putative glutathione S-transferase, could be unambiguously assigned to the chloroplast. Several others （＇potential cp proteins＇） were found to be imported into chloroplasts in vitro, but failed to localize to the organelle when RFP was fused to their C-terminal ends. Extrapolations suggest that the fraction of cp proteins that enter the inner compartments of the organelle, although they lack a cTP, might be as large as 11.4% of the total cp proteome. Our data also support the idea that cytosolic proteins that associate with the cp outer membrane might account for false positive cp proteins obtained in earlier studies.

关键词： Chloroplast biology mitochondria organelle biogenesis/function protein targeting.

Ycf1/Tic214 is Not Essential for the Accumulation of Plastid Proteins

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Molecular Plant 2017年第1期10卷 219-221页

作者： Bettina B?lter Jürgen Soll department Biologie i-botanik Ludwig-Maximilians-Universit?t Gro?hadernerstra?e 2-4 82152 Planegg-Martinsried Germany

Dear Editor,Protein import into chloroplasts is an essential process for plant survival. Numerous studies in recent years have revealed many facts about the components involved and mechanistic features, but have also turned up some controversies. Especially the composition and function of the Tic （translocon at the inner enve- lope of chloroplasts） complex（es） are vividly debated. Whereas the importance of one central Tic component,

关键词： Essential Accumulation Proteins

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

Alb4 of Arabidopsis Promotes Assembly and Stabilization of a Non Chlorophyll-Binding Photosynthetic Complex, the CF1CF0-ATP Synthase

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Molecular Plant 2009年第6期2卷 1410-1424页

作者： Monique Benz Thomas Bals irene Luise Gugel Markus Piotrowski Andreas Kuhn Danja Schunemann Jurgen Soll Elisabeth Ankele department of Biologie i-botanik Biozentrum Ludwig-Maximillians-University Munich Grosshadernerstrasse 2-4 D-82152 Planegg-Martinsried Germany Munich Centre for integrated Protein Science CiPSM Ludwig-Maximillians-University Munich Feodor-Lynen-Strasse 25 D-81377 Munich Germany department of Plant Physiology Ruhr-University Bochum D-44780 Bochum Germany department of Microbiology University of Hohenheim Garbenstrasse 30 D-70599 Hohenheim Germany

All members of the YidC/Oxal/Alb3 protein family are evolutionarily conserved and appear to function in membrane protein integration and protein complex stabilization. Here, we report on a second thylakoidal isoform of Alb3, named Alb4. Analysis of Arabidopsis knockout mutant lines shows that Aib4 is required in assembly and/or stability of the CF1CF0-ATP synthase （ATPase）. alb4 mutant lines not only have reduced steady-state levels of ATPase subunits, but also their assembly into high-molecular-mass complexes is altered, leading to a reduction of ATP synthesis in the mutants. Moreover, we show that Alb4 but not Aib3 physically interacts with the subunits CF1β and CF0ll. Summarizing, the data indicate that Aib4 functions to stabilize or promote assembly of CF1 during its attachment to the membrane-embedded CF0 part.

关键词： Alb4 Alb3 Arabidopsis thylakoid membrane protein insertion protein complex assembly CF1CF0-ATP synthase.

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

Update on Chloroplast Research： New Tools, New Topics, and New Trends

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Molecular Plant 2011年第1期4卷 1-16页

作者： Ute Armbruster Paolo Pesaresi Mathias Pribil Alexander Hertle Dario Leister Lehrstuhl for Molekularbiologie der Pflanzen (botanik) Department Biologie I Ludwig-Maximilians-Universitat M6nchen Groβhaderner Str. D-82152 Planegg-Martinsried Germany b Dipartimento di Scienze Biomolecolari e Biotecnologie Universita degli studi di Milano 1-20133 Milano Italy

Chloroplasts, the green differentiation form of plastids, are the sites of photosynthesis and other important plant functions. Genetic and genomic technologies have greatly boosted the rate of discovery and functional character- ization of chloroplast proteins during the past decade. indeed, data obtained using high-throughput methodologies, in particular proteomics and transcriptomics, are now routinely used to assign functions to chloroplast proteins. Our knowl- edge of many chloroplast processes, notably photosynthesis and photorespiration, has reached such an advanced state that biotechnological approaches to crop improvement now seem feasible. Meanwhile, efforts to identify the entire com- plement of chloroplast proteins and their interactions are progressing rapidly, making the organelle a prime target for systems biology research in plants.

关键词： Chloroplast biology genetics genomics molecular biology proteomics transcriptome analysis

A large-scale phylogenetic revision of Roccellaceae(Arthoniales)reveals eight new genera

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Fungal Diversity 2015年第1期70卷 31-53页

作者： Damien Ertz Anders Tehler Martin irestedt Andreas Frisch Göran Thor Pieter van den Boom Jardin Botanique National de Belgique Département Bryophytes-ThallophytesDomaine de Bouchout1860 MeiseBelgium Naturhistoriska riksmuseet Enheten för botanikBox 5000710405 StockholmSweden Naturhistoriska riksmuseet Enheten för bioinformatik och genetikBox 5000710405 StockholmSweden department of Botany National Museum of Nature and Science4-1-1 AmakuboTsukuba-CityIbarakiJapan 305-0005 department of Ecology Swedish University of Agricultural SciencesP.O.Box 704475007 UppsalaSweden Arafura 16 5691 JA SonThe Netherlands

A two-locus phylogenetic hypothesis of the family Roccellaceae is presented based on data from the nuclear ribosomal large subunit(nucLSU)and the second largest subunit of RNA polymerase ii(RPB2).This analysis includes 341 sequences(166 newly generated)and 180 specimens representing about 114 *** genera Lecanactis,Roccellina,Schismatomma and Sigridea were found to be paraphyletic/*** order to make these groups monophyletic,the new genera Crocellina,Diromma,Gyrographa,Gyronactis,Ocellomma,Pseudoschismatomma,Psoronactis and Vigneronia are *** genus Sagenidium is placed in synonymy with *** new species Enterographa incognita,Gyronactis asiatica and Lecanactis submollis are *** species of Opegrapha are transferred to the *** morphs are recorded for the first time in the genus Syncesia.

关键词： Arthoniomycetes Lichenized fungi Molecular phylogenetics New genera Taxonomy

A Short Amino-Terminal Part of Arabidopsis Phytochrome A induces Constitutive Photomorphogenic Response

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Molecular Plant 2012年第3期5卷 629-641页

作者： Andra＇s Viczia＇n E＇va Ada＇m iris Wolf Ja＇nos Bindics Stefan Kircher Marc Heijde Roman Ulm Eberhard Scha＇fer Ferenc Nagy institute of Plant Biology Biological Research Centre of the Hungarian Academy of Sciences Temesva＇ri krt. 62. H-6726 Szeged Hungary Biologie ii/institut for botanik University of Freiburg Freiburg Dermany department of Botany and Plant Biology University of Geneva Geneva Switzerland Centre for Biological Signalling Studies （BiOSS） University of Freiburg Freiburg Germany School of Biological Sciences University of Edinburgh Edinburgh UK

Phytochrome A （phyA） is the dominant photoreceptor of far-red light sensing in Arabidopsis thaliana, phyA accumulates at high levels in the cytoplasm of etiolated seedlings, and light-induced phyA signaling is mediated by a com- plex regulatory network. This includes light- and FHY1/FHL protein-dependent translocation of native phyA into the nucleus in vivo. it has also been shown that a short N-terminal fragment of phyA （PHYA406） is sufficient to phenocopy this highly regulated cellular process in vitro. To test the biological activity of this N-terminal fragment of phyA in planta, we produced transgenic phyA-201 plants expressing the PHYA406-YFP （YELLOW FLUORESCENT PROTEiN）-DD, PHYA406- YFP-DD-NLS （nuclear localization signal）, and PHYA406-YFP-DD-NES （nuclear export signal） fusion proteins. Here, we report that PHYA406-YFP-DD is imported into the nucleus and this process is partially light-dependent whereas PHYA406-YFP-DD-NLS and PHYA406-YFP-DD-NES display the expected constitutive localization patterns. Our results show that these truncated phyA proteins are light-stable, they trigger a constitutive photomorphogenic-like response when localized in the nuclei, and neither of them induces proper phyA signaling. We demonstrate that in vitro and in vivo PHYA406 Pfr and Pr bind COP1, a general repressor of photomorphogenesis, and co-localize with it in nuclear bodies. Thus, we conclude that, in planta, the truncated PHYA406 proteins inactivate COP1 in the nuclei in a light-independent fashion.

关键词： Arabidopsis cop phenotype phytochrome A COP1

Meta-Analysis of Retrograde Signaling in Arabidopsis thaliana Reveals a Core Module of Genes Embedded in Complex Cellular Signaling Networks

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Molecular Plant 2014年第7期7卷 1167-1190页

作者： Christine GlaiBer Georg Haberer iris Finkemeier Thomas Pfannschmidt Tatjana Kleine Dario Leister Kari-Josef Dietz Rainer Erich Haiusler Bernhard Grimm Klaus Franz Xaver Mayer Helmholtz Zentrum Munchen German Research Center for Environmental Health Institute of Bioinformatics and Systems Biology(IBIS) Ingolstadter Landstr. 1 D-85764 Neuherberg Germany Biozentrum der LMU Munchen Department of Biologie I-Botanik Groβhaderner Str. 2-4 D-82152 Planegg-Martinsried Germany Friedrich-Schiller-Universitat Jena Institut fur AIIgemeine Botanik und Pflanzenphysiologie Dornburger Str. 159 D-07743 JenaGermany Laboratoire de Physiologie Cellulaire Vegetale (LPCV) CEA/CNRS/UJF iRTSV CEA Grenoble 17 rue des Martyrs 38054 Grenoble cedex 9France Biochemistry and Physiology of Plants Faculty of Biology Bielefeld University Universit~tsstrai3e 25 D-33615 Bielefeld Germany University of Cologne Botanical Institute Cologne Biocenter Zu1picher Str. 47B D-50674 Cologne Germany Humboldt-Universitat zu Berlin Institut fur Biologie AG Pflanzenphysiologie Philippstrasse 13 D-10115 Berlin Germany

Plastid-to-nucleus signaling is essential for the coordination and adjustment of cellular metabolism in response to environmental and developmental cues of plant cells. A variety of operational retrograde signaling path- ways have been described that are thought to be triggered by reactive oxygen species, photosynthesis redox imbalance, tetrapyrrole intermediates, and other metabolic traits. Here we report a meta-analysis based on transcriptome and pro- tein interaction data. Comparing the output of these pathways reveals the commonalities and peculiarities stimulated by six different sources impinging on operational retrograde signaling. Our study provides novel insights into the interplay of these pathways, supporting the existence of an as-yet unknown core response module of genes being regulated under all conditions tested. Our analysis further highlights affiliated regulatory cis-elements and classifies abscisic acid and auxin-based signaling as secondary components involved in the response cascades following a plastidial signal. Our study provides a global analysis of structure and interfaces of different pathways involved in plastid-to-nucleus signaling and a new view on this complex cellular communication network.

关键词： retrograde signaling meta-analysis abscisic acid auxin.