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Institut de Biologie StructuraleGrenoble / France

Contact person(s) related to this article / NEUMANN Emmanuelle / SCHOEHN Guy

Presentation of the Methods and Electron Microscopy group

Head of group : Guy SCHOEHN

The group is divided into 3 teams and is also in charge of the EM platform :

Team Guy Schoehn : Macromolecular assemblie’s structure by Electron Microscopy (EM)
Members : Benoit Arragain, Grégory Effantin, Leandro Estrozi, Daphna Fenel, Benoit Gallet, Dominique Housset, Romain Linares, Wai-Li Ling, Hélène Malet, Christine Moriscot, Emmanuelle Neumann, Guy Schoehn

Team Jean-Luc Pellequer  : AFM
Members : Shu-wen Wendy Chen, Jean-Luc Pellequer, Jean-Marie Teulon

Team Pascal Fender  : Adenovirology
Members : Solène Besson, Marie-Claire Dagher, Pascal Fender, Marc André Hograindleur, Emilie Stermann

EM Platform  : Quality Control, Training, Access to equipments, Cellular Electron Microscopy
Members : Grégory Effantin, Daphna Fenel, Benoit Gallet, Christine Moriscot, Emmanuelle Neumann, Guy Schoehn

Aims of the group

• Structure determination of biological macromolecular complexes by EM.
• Provide expertise for EM in single particle analysis, tomography, cellular imaging, and the accompanying computing facilities for both EM and AFM.
• Participate in national nanometrology surveys around the characterization of NPs of biological interest (size and shape).
• Develop a bionanomechanical expertise for probing stiffness of single cells or tissues.
• Use structural virology approaches to study the internalization of adenoviruses and the replication of Bunyaviridae and other negative strained RNA viruses.

Projects

- EM Platform developments and activity (Team Schoehn). The PF aims at collaborating with other IBS teams and to offer either its expertise or trainings to allow scientists to become independent in EM. 
- Bionanomechanics in plant roots (Team Pellequer). A European Training Network grant has been awarded to a consortium of researchers mainly originating from our previous COST Action TD1002 ; the AFM team coordinates the training working package. Our project aims at studying the mechanical response of root tissues during abiotic stress for understanding the STOP1-LPR1 multi-stress signaling pathways in Arabidopsis plants.
- Development of a patented polyvalent vaccine platform (Team Fender). The proof of concept of the ADDomer technology has been demonstrated using Chikungunya virus neutralizing epitopes. Our future goal is to extend this result in the field of immuno-oncology.
- Methodological developments (Team Schoehn & Pellequer). The group will pursue its long-term activity in method developments such as image processing (both EM and AFM) or 3D electron diffraction.
- Structural virology (Team Fender & Schoehn). Since the structure of DSG2 in interaction with the Ad3 fiber has just been solved by cryo-EM, other Adenovirus serotypes involved in severe diseases and/or in gene therapy trials (ex : Ad7, Ad11) will be investigated using EM. We also aim to determine the structure of the Hantaan virus polymerase in different functional states to derive a 3D film of the conformational changes of this enzyme in action. We will perform a structural analysis of native RNP in viruses (by tomography) and during cellular infection (cellular ME), to provide an integrated view of replication/transcription processes. Work on measles and mumps viruses is also continuing.

Keywords

Structural biological projects :
- Structure of macromolecular assemblies
- Virus (bacteriophages and negative stranded RNA viruses) life cycle
- Adenovirus life cycle and therapeutic applications
- Bionanomechanics of cell walls of plant roots by AFM
- Nanoparticle metrology and nanotoxicology
- Large biomolecules reconstruction using AFM topography

Methods :
- Electron microscopy (negative staining, cryoEM, cryo-tomography, micro-electron diffraction)
- Image analysis (EM and AFM)
- Protein expression (Baculovirus)
- Biochemistry
- Protein-protein interactions (SPR ; BLI)
- Flow cytometry

New Major References

Vassal-Stermann E, Effantin G, Zubieta C, Burmeister W, Iseni F, Wang H, Lieber A, Schoehn G, Fender P.(2019). CryoEM structure of adenovirus type 3 fibre with desmoglein 2 shows an unusual mode of receptor engagement. Nat Commun. Mar 12 ;10(1):1181. doi : 10.1038/s41467-019-09220-y.

- Polovinkin L, Hassaine G, Perot J, Neumann E, Jensen AA, Lefebvre SN, Corringer PJ, Neyton J, Chipot C, Dehez F, Schoehn G, Nury H. (2018). Conformational transitions of the serotonin 5-HT3 receptor. Nature. 563(7730):275-279.

- Da Silveira Tomé C, Foucher AE, Jault JM, Housset D. (2018) High concentrations of GTP induce conformational changes in the essential bacterial GTPase EngA and enhance its binding to the ribosome.The FEBS Journal 285, 1, pp.160-177.

- Flori S, Jouneau PH, Gallet B, Estrozi LF, Moriscot C, Schoehn G, Finazzi G, Falconet D. (2018). Imaging Plastids in 2D and 3D : Confocal and Electron Microscopy. Methods Mol Biol. 1829:113-122.

- Vassal-Stermann E, Mottet M, Ducournau C, Iseni F, Vragniau C, Wang H, Zubieta C, Lieber A, Fender P. (2018). Mapping of Adenovirus of serotype 3 fibre interaction to desmoglein 2 revealed a novel ‘non-classical’ mechanism of viral receptor engagement. Sci Rep. 8 (1), pp.8381.

- Belime A, Thielens NM, Gravel E, Frachet P, Ancelet S, Tacnet P, Caneiro C, Chuprin J, Gaboriaud C, Schoehn G, Doris E, Ling WL. (2018). Recognition protein C1q of innate immunity agglutinates nanodiamonds without activating complement. Nanomedicine S1549-9634(18)30528-8.

- Pellegrini E, Desfosses A, Wallmann A, Schulze WM, Rehbein K, Mas P, Signor L, Gaudon S, Zenkeviciute G, Hons M, Malet H, Gutsche I, Sachse C, Schoehn G, Oschkinat H, Cusack S.(2018). RIP2 filament formation is required for NOD2 dependent NF-κB signalling. Nat Commun. 9(1):4043.

- El-Khatib M, Nasrallah C, Lopes J, Tran QT, Tetreau G, Basbous H, Fenel D, Gallet B, Lethier M, Bolla JM, Pagès JM, Vivaudou M, Weik M, Winterhalter M, Colletier JP. (2018). Porin self-association enables cell-to-cell contact in Providencia stuartii floating communities. Proc Natl Acad Sci U S A. 115(10):E2220-E2228.

- Neumann E, Farias Estrozi L, Effantin G, Breyton C, Schoehn G. (2017). The resolution revolution in cryo-electron microscopy médecine/sciences, EDP Sciences 33(12):1111-1117.

- Bonnet J, Durmort C, Jacq M, Mortier-Barrière I, Campo N, VanNieuwenhze MS, Brun YV, Arthaud C, Gallet B, Moriscot C, Morlot C, Vernet T, Di Guilmi AM. (2017). Peptidoglycan O-acetylation is functionally related to cell wall biosynthesis and cell division in Streptococcus pneumoniae. Mol Microbiol. 106(5):832-846.

- Arlaud, G.J., Gaboriaud, C., Ling, W.L., Thielens, N. (2017). Structure of the C1 complex of complement. Proceedings of the National Academy of Sciences 114, E5766-5767.

- Rodrigues CD, Henry X, Neumann E, Kurauskas V, Bellard L, Fichou Y, Schanda P, Schoehn G, Rudner DZ, Morlot C. (2016). A ring-shaped conduit connects the mother cell and forespore during sporulation in Bacillus subtilis. Proc Natl Acad Sci U S A. 113(41):11585-11590.

- Kandiah E, Carriel D, Perard J, Malet H, Bacia M, Liu K, Chan SW, Houry WA, Ollagnier de Choudens S, Elsen S, Gutsche I. (2016). Structural insights into the Escherichia coli lysine decarboxylases and molecular determinants of interaction with the AAA+ ATPase RavA. Sci Rep. 6:24601.

- Costa L, Andriatis A, Brennich M, Teulon JM, Chen SW, Pellequer JL, Round A. (2016). Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules. BMC Struct Biol. 16(1):18.

- Gutsche I, Desfosses A, Effantin G, Ling WL, Haupt M, Ruigrok R, Sachse C, Schoehn G. Near-atomic cryo-EM structure of the helical measles virus nucleocapsid. Science (2015) 348 (6235):704-7.

The list of all the publications of the group is available here.