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Our group is interested in the cell biology of complex DNA viruses. How does a virus get access to a cell, disassemble to uncover its genome for replication, and how is - within a couple of hours - the viral program switched to virus assembly and egress?
Research summary
HSV1 enters cells by fusion of the viral envelope with the plasma membrane. The incoming viral capsids are subsequently propeled along microtubules (MTs) to the nucleus using cytosolic dynein, a minus-end directed motor ATPase. They are ultimately targeted via importin beta to the nuclear pore where the viral genome is uncoated for viral transcription and replication in the nucleoplasm. During virus assembly and egress, cytosolic capsids are presumably also transported along MTs and this time possibly use kinesins, MT plus-end directed motor ATPases.
We have developed several in vivo and in vitro motility assays to analyse the cell biology of HSV1 and particularly the MT-mediated capsid transport by digital video microscopy. Using these assays, we characterize cytosolic host factors as well as structural viral proteinsimportant in HSV1 entry and egress. Besides wild type we use a mutant virus that carries instead of the small capsid protein VP26 a fluorescent protein GFP-VP26. Individual GFP-VP26 labelled capsids entering living cells can be analysed with reasonable time resolution by digital time-lapse video microscopy. Using YFP-tubulin expressing cells, we are analysing directly the role of dynein and the dynein cofactor dynactin in cytosolic capsid transport by overexpressing and microinjecting dominant negative subunits of dynein and dynactin. In addition, this system is used to analyse whether capsids are transported from the microtubule organising centre (MTOC) further to the nucleus, and whether this requires kinesins. To dissect the interaction between the viral capsids and the MTs biochemically, we have developed in vitro binding assays. The simplest consists of isolated viral capsids and a cytosolic extract. When the cytosol-capsid mixture is incubated with MTs, capsids can bind to the MTs. Recombinant candidate proteins, peptides or antibodies against cytosolic as well as viral proteins are then tested for their ability to interfere with motor or MT binding. These experiments characterize the molecular interface between the viral capsid and the motor dynein. The ultimate goal of our research is to understand the molecular determinants of cytosolic viral capsid transport. Along these lines, we have developed an in vitro motility assay containing isolated GFP-VP26-capsids, Cy3-labelled MTs, and cytosol. By digital video microscopy, we observe individual GFP-VP26-capsids moving unidirectionally and with constant speed along MTs. Using different cytosol preparations, we determine whether herpes capsids use indeed dynein for MT transport, and try to dissect the stimulatory function of the dynein cofactor dynactin. Using a functional proteomics approach, we mutate several structural viral proteins. We are mainly interested in tegument proteins localized between the viral envelope and the capsid, since we believe that they include putative viral dynein and kinesin receptors. Using a recombinant BAC-vector (bacterial artificial chromosome) encoding the entire HSV1 genome, we will generate specific mutants of HSV1 that will be tested for their ability to move along MTs as well as for the directionality of such transport. In contrast to other cargo transported by MTs, the known protein composition of this one can be manipulated in a controlled manner. Dependent on the stage of the viral life cycle and the cellular context, herpes virus capsids move to the cell centre or periphery by being transported to the MT minus- or plus-ends. We will use our system to identify putative motor receptors and to analyse the factors controlling the directionality of MT transport and the activation of different MT motors. The in vitro assays that we have developed provide tools to analyse other human pathogenic herpes viruses for which so far no good cell culture models have been established such as cytomegalovirus or Kaposi sarcoma-associated herpesvirus. Our studies will hopefully in the long run identify viral-host interactions suitable for drug discovery and new antiviral therapy. Teaching
Virologie für Chemiker, Biochemiker und Biologen
Praktikum I "Virusnachweis" im Sommersemester
Praktikm II "Molekulare und Zelluläre Virologie" im Wintersemester
Biochemisches Praktikum für Fortgeschrittene II für Biochemiker Biologisches Praktikum für Biologen DFG-Graduiertenkolleg 745: Mukosale Erreger-Wirt-Interaktionen MHH-Md/PhD-Programm ZIB - Zentrum für Infektionsbiologie <-- back to top Team Team leader
HD Dr. rer. nat. Beate Sodeik
tel. +49 (511) 532 2846 | fax. +49 (511) 532 2736 sodeik.beate@mh-hannover.de
Dr. Sodeik studied biology in Bonn (1982-88), and obtained a diploma in cell biology analyzing autophagy in the single cell animal Amoeba proteus. She then trained as a DAAD fellow (German Academic Exchange Service; 1988/89) at Columbia University Medical School, College of Physicians & Surgeons, New York, USA working on integrin receptors of human neutrophils. For her PhD project in cell biology (1989-93), she moved to the European Molecular Biology Laboratory (EMBL) in Heidelberg, and initiated a project on the assembly of vaccinia virus in the laboratory of Dr. Gareth Griffiths. She joined the group of Prof. Ari Helenius at Yale University, New Haven, CT, USA as an EMBO postdoctoral fellow and later associate, to set up the analysis of virus-host interactions during the early phase of herpes simplex virus infection (1993-97). From 1997 until 2002 she was a senior scientist and team leader at Hannover Medical School in the Department of Biochemistry. After her postdoctoral thesis in biochemistry ("Habilitation"), she was appointed as assistant professor in the Department of Virology at Hannover Medical School in 2002.
Research staff
Anne Binz, Research Assistant
+49 (511) 532 2845 | Binz.Anne@MH-hannover.deKatinka Döhner, Dipl. Biochem.; PhD
+49 (511) 532 2845 | Katinka.Doehner@stud.uni-hannover.deMojgan Fathollahy, Research assistant
+49 (511) 532 2845 | Fathollahy.Mojgan@MH-Hannover.deJanus, Jessica; Dipl. Biol.
+49 (511) 532 2845 | Janus.Jessica@mh-hannover.deClaus-Henning Nagel, Dipl. Biochem.
+49 (511) 532 2845 | Claus-Henning.Nagel@stud.uni-hannover.deMichaela Nygårdas; Visiting PhD student from Turku, Finland
+49 (511) 532 2845 | Nygardas.Michaela@MH-Hannover.deKlaas Mulder; Visiting Master student from Amsterdam, The Netherlands
+49 (511) 532 2845 | klaasmulder2002@hotmail.comUte Prank, Research assistant
+49 (511) 532 2845 | Prank.Ute@MH-hannover.deKerstin Radtke; Dipl. Biochem.
+49 (511) 532 2845 | Radtke.Kerstin@MH-hannover.deKathrin Rode; Dipl. Biochem.
+49 (511) 532 2845 | Kathrin.Rode@stud.uni-hannover.deJulia Schipke; Dipl. Biol.
+49 (511) 532 2845 | Schipke.Julia@MH-hannover.deKristina Theusner; Master in Biology
+49 (511) 532 2845 | Theusner.Kristina@MH-hannover.deFormer members of the group
André Wolfstein, Dipl. Ing., Dr. rer. nat. (1998-2006) Simone Schmidt; veterinary doctor (2001-2003) Sabrina Marozin; veterinary doctor (2001-2002) Tanja Strive; Dr. rer. nat. (2001-2002) <-- back to top The German Research Council (Deutsche Forschungsgemeinschaft) funds our research with a project grant (So403/1), a national research co-operative on Molecular Motors (DFG-Schwerpunktprogramm; So403/2), a national research cooperative on Membrane budding and its exploitation by viruses (DFG-Schwerpunkt-programm; So403/3) and the joint graduate research program of the Hannover Medical and Veterinary Schools (Graduiertenkolleg 745: Mucosal host-pathogen interactions). Moreover, we obtain financial support from the EU in the NEST adventure program on Axonuclear communication in health and disease, from the Minister for Science and Culture (Volkswagen Stiftung) in collaboration with the Technion University (Israel), from the DAAD in collaboration with the Universities of Helsinki and Turku, both Finland, and the Centre of Infection Biology at the Hannover Medical School (MHH - ZIB).
<-- back to top Primary publications in peer-reviewed journals
Döhner, Radtke, Schmidt & Sodeik (2006). The eclipse phase of Herpes Simplex Virus type 1 infection: Efficient dynein-mediated nuclear capsid targeting without the small capsid protein VP26. Journal of Virology, in press.
Köllner, Sodeik, Schreek, Heyn, von Neuhoff, Germeshausen, Zeidler, Krüger, Schlegelberger, Welte, Beger (2006). Mutations in neutrophil elastase causing congenital neutropenia lead to cytoplasmic protein accumulation and induction of the unfolded protein response. Blood, in press. Wolfstein, Nagel, Radtke, Döhner, Allan & Sodeik (2006). The inner tegument promotes herpes simplex virus capsid motility along microtubules in vitro. Traffic 7:227-237. Hass, Siegert, Schürmann, Sodeik & Wolfes (2004). The c-Myb protein interacts with Rcd-1, a component of the CCR4 transcription mediator complex. Biochemistry 25:8152-59. Marozin, Prank & Sodeik (2004). Herpes simplex virus type 1 infection of polarized epithelial cells requires microtubules and access to receptors present at cell-cell contact sites. Journal of General Virology 85: 775-786. H Mabit, MY Nakano, U Prank, B Saam, K Döhner, B Sodeik & U Greber (2002). "Intact microtubules support adenovirus and herpes simplex virus infection."Journal of Virology 76:9962-9971. K Döhner, A Wolfstein, U Prank, C Echeverri, D Dujardin, R Vallee, & B Sodeik (2002). "Function of dynein and dynactin in herpes simplex virus capsid transport."Molecular Biology of the Cell 13:2795-2809. PM Ojala (corresponding first author), B Sodeik, MW Ebersold, U. Kutay, A Helenius (2000). "Herpes simplex virus 1 capsid binding to nuclei and genome uncoating at the nuclear pore complex in vitro."Molecular & Cellular Biology 20:4922-4931. M Kann (corresponding first author), B Sodeik, A Valchou, WH Gerlich, A. Helenius (1999). "Phosphorylation-dependent binding of hepatitis B core particles to the nuclear pore complex." Journal of Cell Biology 145:45-55. A Alconada, U Bauer, B Sodeik, B Hoflack (1999). "Intracellular traffic of herpes simplex virus glycoprotein gE: Characterization of the sorting signals required for its trans-Golgi network localization." Journal of Virology 73:377-387. M Ericsson, B Sodeik, J Krijnse-Locker, G Griffiths (1997). "Reconstitution of vaccinia virus assembly in vitro after release from the rifampicin block."Virology 235:218-227. B Sodeik (corresponding first author), MW Ebersold, A Helenius (1997). "Microtubule mediated transport of incoming herpes simplex virus I capsids to the nucleus." Journal of Cell Biology 136:1007-1021. S Cudmore, R Blasco, R Vincentelli, M Esteban, B Sodeik, G Griffiths, J Krijnse-Locker (1996). "A vaccinia virus core protein, p39, is membrane associated." Journal of Virology 70:6909-6921. B Sodeik, S Cudmore, M Ericsson, M Esteban, EG Niles, G Griffiths (1995). "Assembly of vaccinia virus: Incorporation of two membrane proteins into the intracellular mature virus." Journal of Virology 69:3560-3574. B Sodeik, G Griffiths, M Ericsson, B Moss, RW Doms (1994). "Assembly of vaccinia virus: Effects of rifampicin on the intracellular distribution of viral protein p65." Journal of Virology 68:1103-1114. M Schmelz, B Sodeik, M Ericsson, EJ Wolffe, H Shida, G Hiller, G Griffiths (1994). "Assembly of vaccinia virus: The second wrapping cisterna is derived from the trans Golgi network." Journal of Virology 68:130-147. B Sodeik, RW Doms, M Ericsson, G Hiller, CE Machamer, W van' t Hof, G van Meer, B Moss, G Griffiths (1993). "Assembly of vaccinia virus: Role of the intermediate compartment between the endoplasmic reticulum and the Golgi Stack." Journal of Cell Biology 121:521-541. JD Loike, B Sodeik, L Cao, S Leucona, JI Weitz, PA Detmers, SD Wright and SC Silverstein (1991). "CD11c/CD18 on neutrophils recognizes a domain at the N terminus of the A? chain of fibrinogen." Proceedings of the National Academy of Sciences USA 88,1044-1048. B Sodeik, K Brix, W Stockem (1989)."Sequestration of microinjected molecular probes from the cytoplasm of Amoeba proteus." European Journal of Protistology 25:75-84. <-- back to top Reviews, Book chapters, comments etc.
Radtke, Döhner & Sodeik (2006). Viral interactions with the cytoskeleton: a hitchhiker's guide to the cell. Cellular Microbiology 8:387-400 (invited review).
Döhner, Nagel & Sodeik (2005). Viral stop-and-go along microtubules - Taking a ride with dynein and kinesins. Trends in Microbiology, 13:320-327 (invited review). Sodeik, Schramm, Suomalainen & Krijnse-Locker (2005). Meeting report: EMBO Workshop - Cell Biology of Virus Infection. Traffic 6:351-356. Döhner & Sodeik (2004). The role of the host cytoskeleton for viral replication. Current Topics in Microbiology and Immunology 285, 67-108 (invited review). Sodeik & Kann (2003). From the entry to the progeny virus - Workshop des Arbeitskreises ZELLBIOLOGIE in der Gesellschaft für Virologie. Zellbiologie aktuell 29:27-28. Sodeik (2002). Unchain my heart, baby let me go - the entry and intracellular transport of HIV. Journal of Cell Biology 159:393-395. Sodeik & Krijnse-Locker (2002). Assembly of vaccinia virus revisited - Double membrane acquisition from a host organelle? Trends in Microbiology 10:15-24 (invited review). Krijnse-Locker, Sodeik, & Suomalainen (2002). Meeting report on the EMBO Workshop - The Cell Biology of Virus Infection. Traffic 3:233-235. Wolfstein & Sodeik (2001). Capsid motility of herpes simplex virus type 1 along microtubles in vitro. Trends in Cell Biology, CD - GFP in Motion 2 - Live Fluorescence Imaging using Green fluorescent protein; compiled by B Ludin & A Matus (invited contribution on live cell imaging). Sodeik (2000). Mechanisms of viral transport in the cytoplasm. Trends in Microbiology 8:419-426 (invited review). Izaurralde, Kann, Pante, Sodeik, & Hohn (1999). EMBO workshop report. Viruses, microorganisms and scientists meet the nuclear pore. EMBO Journal 18:289-296. Griffiths & Sodeik (1992). The biogenesis of vaccinia virus. In: ed. J.A.F. Op den Kamp, NATO ASI Series. 63:365-374, Springer-Verlag, Berlin. |