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Arbeitsgruppe: Prof. Danja Schünemann

 

Chloroplasts contain approximately 3500 proteins, but only a small fraction of these proteins (approximately 100) are encoded in the chloroplast genome. Most chloroplast proteins are encoded in the nucleus and have to be imported post-translationally across the translocon of the inner and outer envelope into the organelle. For the translocation of proteins from the stroma to the thylakoid membrane or across the membrane into the thylakoid lumen four different pathways have been described so far: the Sec-pathway and the Tat-pathway are mainly involved in the transport of lumenal proteins, whereas the spontaneous-pathway and the cpSRP-pathway are used for the insertion of integral membrane proteins into the thylakoid membrane. Current research projects of our group are the detailled structure-function analysis of proteins that are known to be involved in the targeting of proteins to the thylakoid membrane, identification of novel components required for protein transport to or across the thylakoid membrane and analysis of the regulation of the cpSRP-dependent protein transport.

 
S-Block / S-Modul

Molekularbiologische und proteinbiochemische Untersuchungen zum plastidären Proteintransport
(SS und WS nach Vereinbarung; 4-6 Wochen ganztägig)
Inhalt:
pChloroplasten besitzen ungefähr 2500 Proteine. Über 95 % dieser Proteine sind im Kern kodiert. Wie erreichen die kernkodierten  Proteine ihre chloroplastidären Bestimmungsorte? Dieses Problem ist kompliziert, da bei der Zielsteuerung der Proteine zum Chloroplasten zwischen sechs Bestimmungsorten unterschieden werden muß (äußere Hüllmembran, Intermembranraum, innere Hüllmembran, Stroma, Thylakoidmembran, Thylakoidlumen). Im Rahmen des Praktikums sollen die Studenten Experimente zu verschiedenen Teilaspekten folgender Fragen durchführen:

  • Welche stromalen Faktoren sind an der spezifischen Erkennung der aus dem Cytosol in den Plastiden importierten Proteinen beteiligt?
  • Wie wird der Transport der Proteine zu den Thylakoidmembranen der Chloroplasten gesteuert?
  • Wie erfolgt der Durchtransport eines Makromoleküls durch eine im Prinzip undurchlässige Membran?

Lernziele:

Es sollen verschiedene molekularbiologische und proteinchemische Techniken erlernt werden (z. B. Synthese von rekombinanten Proteinen durch Überexpression in Bakterien und in vitro Translation, Herstellung von Deletions- und Punktmutationskonstrukten verschiedener Proteine, Hefe two hybrid Systeme zur Analyse von Protein-Protein-Interaktionen, Proteinauftrennung durch FPLC). In begleitenden Veranstaltungen in Form von Seminaren und Vorträgen sollen die Studenten die Darstellung und die Bewertung von experimentellen Daten üben.
 

Veröffentlichungen

2015

  • Walter B, Hristou A, Nowaczyk MM, Schünemann D (2015) In vitro
    reconstitution of cotranslational D1 insertion reveals a role of the
    cpSec/Alb3 translocase and Vipp1 in photosystem II biogenesis.
    Biochem J. 2015 Mar 24. [Epub ahead of print]  Weblink Weblink
  • Dünschede B, Träger C, Schröder CV, Ziehe D, Walter B, Funke S, Hofmann
    E, Schünemann D (2015) Chloroplast SRP54 was Recruited for
    Posttranslational Protein Transport via Complex Formation with
    Chloroplast SRP43 during Land Plant Evolution. Weblink Weblink
    J Biol Chem. 2015 Apr 1. pii: jbc.M114.597922. [Epub ahead of print]
  • Walter B, Pieta T and Schünemann D (2015) Arabidopsis thaliana mutants
    lacking cpFtsY or cpSRP54 exhibit different defects in photosystem II
    repair.
    Front. Plant Sci. 6:250.  Weblink

2014

  • Schönberg A, Bergner E, Helm S, Dünschede B, Schünemann D, Schutkowski M; Baginsky S (2014) The peptide chip “ChloroPhos1.0” identifies new phosphorylation targets of plastid casein kinase II (pCKII) in Arabidopsis thaliana.
    PLoS One 2014 Oct 8;9(10):e108344 Weblink Weblink
  • Rühle T, Razeghi JA, Vamvaka E, Viola S, Gandini C, Kleine T, Schünemann D, Barbato R, Jahns P, Leister D (2014) The Arabidopsis protein CONSERVED ONLY IN THE GREEN LINEAGE160 promotes the assembly of the membranous part of the chloroplast ATP synthase.
    Plant Physiol 165: 207-226 Weblink Weblink
  • Schneider A, Steinberger I, Strissel H, Kunz HH, Manavskia N, Meurer J, Burkhard G, Jarzombski S, Schünemann D, Geimer S, Flügge UI and Leister D (2014) The Arabidopsis TELLURITE RESISTANCE C Protein together with ALB3 is involved in Photosystem II Protein Synthesis.
    The Plant Journal 78: 344-356 Weblink Weblink

2013

  • Rosenblad MA, Träger C, Schünemann D. (2013) Structural diversity of signal recognition particle RNAs in plastids.
    Plant Signal Behav. 2013 Oct 30;9(1). pii: e26848. [Epub ahead of print] Weblink

2012

  • Träger C., Rosenblad M., Ziehe D., Garcia-Petit C., Schrader L., Kock K., Richter C., Klinkert B., Narberhaus F., Herrmann C., Hofmann E., Aronsson H. and Schünemann D. (2012) Evolution from the prokaryotic to the higher plant chloroplast signal recognition particle: the signal recognition particle RNA is conserved in plastids of a wide range of photosynthetic organisms.
    Plant Cell 24:4819-4836 Weblink Weblink

2011

  • Dünschede B., Bals T., Funke S., Schünemann D. (2011) Interaction studies between the chloroplast signal recognition particle subunit cpSRP43 and the full-length translocase Alb3 reveal a membrane-embedded binding region in Alb3 protein.
    J. Biol. Chem., 286(40):35187-35195 Weblink Weblink
  • Bals T., Schünemann D. (2011) Isolation of Arabidopsis thylakoid membranes and their use for in vitro protein insertion or transport assays.
    Methods Mol. Biol., 774: 321-338 Weblink
  • Bals T., Schünemann D. (2011) The yeast split-ubiquitin system for the analysis of membrane protein interactions.
    Endocytobiosis and Cell Research; 43-51 Weblink
  • Dünschede B., Schünemann D. (2011) The role of Alb3 and its homologs in the insertion and assembly of thylakoid membrane proteins.
    Endocytobiosis and Cell Research: 19-251 Weblink

2010

  • Armbruster U., Zühlke J., Rengstl B., Kreller R., Makarenko E., Rühle T.,
    Schünemann D., Jahns P., Weisshaar B., Nickelsen J., and Leister D. (2010)
    The Arabidopsis Thylakoid Protein PAM68 Is Required for Efficient D1 Biogenesis and Photosystem II Assembly.
    Plant Cell, 22: 3439-34601 Weblink
  • Bals T., Dünschede B., Funke S., Schünemann D. (2010) Interplay between the cpSRP pathway components, the substrate LHCP and the translocase Alb3: an in vivo and in vitro study.
    Febs Lett. 584: 4138-41441 Weblink
  • Richter C., Bals T., Schünemann D. (2010) Component interactions, regulation and mechanisms of chloroplast SRP-dependent protein transport.
    Eur J Cell Biol. 89: 965-9731 Weblink
  • Ellinger D., Stingl N., Kubigsteltig I., Bals T., Jünger M., Pollmann S., Berger S., Schünemann D., Müller M.J. (2010) DONGLE and DEFECTIVE IN ANTHER DEHISCENCE1 lipases are not essential for wound- and pathogen-induced jasmonate biosynthesis: redundant lipases contribute to jasmonate formation.
    Plant Physiol. 153: 114-121 Weblink

2009

  • Benz M., Bals T., Gügel I.L., Piotrowski M., Kuhn A., Schünemann D., Soll J.,
    Ankele E. (2009) Alb4 of Arabidopsis promotes assembly and stabilisation of a none chlorophyll-binding photosynthetic complex, the CF1CF0-ATP synthase. Mol. Plant, 2: 1410-1424 Weblink
  • Schottkowski M., Gkalympoudis S., Tzekova N., Stelljes C., Schünemann D., Ankele E., Nickelsen J. (2009) Interaction of the periplasmic PratA factor and the PsbA (D1) protein during biogenesis of photosystem II in Synechocystis sp. PCC 6803.
    J. Biol. Chem., 284: 1813-18191 Weblink

2008

  • Richter C.V., Träger C., Schünemann D. (2008) Evolutionary substitution of two amino acids in chloroplast SRP54 of higher plants cause its inability to bind SRP RNA.
    FEBS Lett., 582: 3223-32291 Weblink
  • Dal Corso G., Pesaresi P., Masiero S., Aseeva E., Schünemann D., Finazzi G., Joliot P., Barbato R., Leister D. (2008) A complex containing PGRL1 and PGR5 is involved in the switch between linear and cyclic electron flow in Arabidopsis.
    Cell, 132: 273-281 Weblink

2007

  • Schünemann D. (2007) Mechanisms of protein import into thylakoids of chloroplasts.
    Biol. Chem., 388: 907-9151 Weblink
  • Henry R., Goforth R., Schünemann D. (2007) The Chloroplast SRP/FtsY and Alb3 in the Integration into the Thylakoid Membrane.
    In: Ross E. Dalbey, Carla Koehler and Fuyuhiko Tamanoi (ed) The Enzymes, Molecular Machines involved in Protein Transport across Cellular Membranes, 25: 493-5211 Weblink

2006

  • Hermkes R., Funke S., Richter C., Kuhlmann J., Schünemann D. (2006) The α-Helix of the second chromodomain of the 43 kDa subunit of the chloroplast signal recognition particle facilitates binding to the 54 kDa subunit.
    FEBS Lett., 580: 3107-31111 Weblink
  • Gerdes L., Bals T., Klostermann E., Karl M., Philippar K., Hünken M., Soll J., Schünemann D. (2006) A second thylakoid membrane localized Alb3/Oxa1/YidC homologue is involved in proper chloroplast biogenesis in Arabidopsis thaliana.
    J. Biol. Chem., 281:16632-16641 Weblink

2005

  • Pasch J.C., Nickelsen J., Schünemann D. (2005) The yeast split-ubiquitin system to study chloroplast membrane protein interactions.
    Appl Microbiol Biotechnol, 69: 440-4471 Weblink
  • Funke S., Knechten T., Ollesch J., Schünemann D. (2005) A unique sequence motif in the 54 kD subunit of the chloroplast signal recognition particle mediates binding to the 43 kD subunit.
    J. Biol. Chem. 280: 8912-89171 Weblink

2004

  • Schünemann D. (2004) Structure and function of the chloroplast signal recognition particle (cpSRP)
    Current Genetics 44: 295-3041 Weblink

2002

  • Klostermann E., Droste gen. Helling I., Carde J.-P., Schünemann D. (2002) The thylakoid membrane protein ALB3 associates with the cpSecY-translocase in Arabidopsis thaliana.
    Biochem. J. 368: 777-7811 Weblink

2001

  • Jonas-Straube E., Hutin C., Hoffman N. E. , Schünemann  D. (2001) Functional analysis of the protein interacting domains of chloroplast SRP43. J. Biol. Chem. 276: 24654-246601 Weblink

1999

  • Tu C., Schünemann D., Hoffman N. E. (1999) Chloroplast FtsY, chloroplast signal recognition particle, and GTP are required to reconstitute the soluble phase of light-harvesting chlorophyll protein transport into thylakoid membranes.
    J. Biol. Chem. 274: 27219-272241 Weblink
  • Schünemann D., Amin P., Hartmann E., Hoffman N. E. (1999) Chloroplast SecY is complexed to SecE and involved in the translocation of the 33-kDa but not the 23-kDa subunit of the oxygen-evolving complex.
    J. Biol. Chem. 274: 12177-121821 Weblink
  • Schünemann D., Amin P., Hoffman N. E. (1999) Functional divergence of the plastid and cytosolic forms of the 54 kD subunit of signal recognition particle.
    Biochem. Biophys. Res. Commun. 254: 253-2581 Weblink
  • Klimyuk V. I., Persello-Cartieaux F., Havaux M., Contard P., Meiherhoff K., Schünemann D., Gouet P., Jones J. D. G., Hoffman N. E., Nussaume L. (1999) A chromodomain protein encoded by the arabidopsis Cao gene is a plant-specific component of the chloroplast signal recognition particle pathway that is involved in LHCP targeting.
    Plant Cell 11: 87-991 Weblink

1998

  • Hoffmann N. E., Schuenemann D., Tu C. J., Amin P., Sy D., Oilgrim M., Gupta S. and Nussaume L (1998) Structure of the chloroplast signal recognition particle and its role in chloroplast biogenesis.
    In Photosynthesis: Mechanisms and Effects, Garab G (ed.), Kluwer Academis Publishers, Pays Bas, Vol IV: 3075-30801 Weblink
  • Schünemann D., Gupta S., Persello-Cartieaux F., Klimyuk V. L., Jones J. D. G., Nussaume L., and Hoffman N. E. (1998) A novel signal recognition particle targets light harvesting proteins to the thylakoid membranes.
    Proc. Natl. Acad. Sci. 95: 10312-103161 Weblink
  • Büker M., Schünemann D., Borchert S. (1998) Enzymic properties and capacities of developing tomato (Lycopersicon esculentum L.) fruit plastids.
    J. Exp. Bot. 49: 681-6911 Weblink

1996

  • Schünemann, D., Schott, K., Borchert, S., Heldt, H. W. (1996) Evidence for the expression of the triosephosphate translocator gene in green and nongreen tissues of tomato and potato.
    Plant Mol. Biol. 31: 101-1111 Weblink

1994

  • Schünemann D., Borchert S. (1994) Specific transport of inorganic phosphate and C3- and C6-sugar-phosphates across the envelope membranes of tomato (Lycopersicon esculentum) leaf-chloroplasts, tomato fruit-chloroplasts and fruit-chromoplasts.
    Bot. Acta 107: 461-4671 
  • Fischer K., Weber A., Brink S., Arbinger B., Schünemann D., Borchert S., Heldt H. W., Popp B., Benz R., Link T. A., Eckerskorn C., Flügge U. I. (1994) Porins from plants: molecular cloning and functional characterization of two new members of the porin family.
    J. Biol. Chem. 269: 25754-257601 Weblink

1993

  • Borchert S., Harborth J., Schünemann D., Hoferichter P., Heldt H.W. (1993) Studies of the enzymatic capacities and transport properties of pea root plastids.
    Plant Physiol. 101: 303-3121 Weblink
  • Schünemann D., Borchert S., Flügge U.-I., Heldt H. W. (1993) ADP/ATP translocator from pea root plastids: comparison with translocators from spinach chloroplasts and pea leaf mitochondria.
    Plant Physiol. 103: 131-1371 Weblink

Kontakt

Prof. Dr. Danja Schünemann
Ruhr-Universität Bochum
AG Molekularbiologie pflanzlicher Organellen

Raum: ND 3/35
Universitätsstr. 150
44780 Bochum

Tel.: 0234-32-24293
E-mail: danja.schuenemann@rub.de
Sprechstunde nach Vereinbarung
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leitung

  • Danja Schünemann
    Prof. Dr. Danja Schünemann
    Ruhr-Universität Bochum
    AG Molekularbiologie
    pflanzlicher Organellen
    Raum: ND 2/67
    Universitätsstraße 150
    44780 Bochum
    Tel.: 0234-32-24293
    E-Mail schicken
    Sprechstunde: Dienstags 9-10 Uhr und nach Vereinbarung
 

Presse

  • Lichtsammler an Ort und Stelle bringen
    RUB-Biologen veröffentlichen neues Modell zum Proteintransport in Pflanzenzellen weiter
  • Auf halbem Weg zwischen Bakterium und Baum
    Wie sich die Protein-Transport-maschinerie in den Chloroplasten höherer Pflanzen entwickelte weiter
 

Mitarbeiter/innen

  • Silke Funke
    Silke Funke
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Beatrix Dünschede
    Dr. Beatrix Dünschede
    Postdoktorandin
    Büro: ND 2/72 (0234/32-28976)
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Dominik Ziehe
    Dominik Ziehe
    Doktorand
    Büro: ND 2/72 (0234/32-28976)
    Labor: ND ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Athina Hristou
    Athina Hristou
    Doktorandin
    Büro: ND 2/72 (0234/32-28976)
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Pascal Radtke
    Pascal Radtke
    Master-Student
    Büro: ND 2/73 (0234/32-28461)
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Saskia Heybrock
    Saskia Heybrock
    Master-Studentin
    Büro: ND 2/73 (0234/32-28461)
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken
  • Mira Zenker
    Mira Zenker
    Bachelor-Studentin
    Büro: ND 2/73 (0234/32-28461)
    Labor: ND 2/71 (0234/32-28467)
    E-Mail schicken