Membrane virology of viral infection
We are interested in studying how viruses interact with cellular membranes and lipids during infection. We use cryo-electron microscopy techniques in conjunction with other imaging methods, such as fluorescence microscopy and imaging mass spectrometry, to structurally characterize influenza A and Ebola virus protein-membrane interactions during entry.
We study how membrane enveloped viruses such as influenza A virus, Ebola virus and SARS-CoV-2 interact with host membranes during their replication. Influenza A virus and Ebola virus form long filamentous virions that must enter the host cell and undergo membrane fusion in order to establish an infection. The mechanism of IAV and Ebola virus disassembly and membrane fusion are not completely understood and have not been structurally analyzed inside the cells. In addition, we are interested how these viruses shape cellular membranes during viral assembly and genome incorporation. We apply a cryo-correlative light and electron microscopy workflow and cryo-electron tomography to provide structural details of pivotal viral replication steps such as membrane fusion, viral genome nucleocapsid organization, and virus assembly.
SARS-CoV-2 assembly in infected cells by in cellulo cryo-electron tomography (Klein, Cortese, Winter, et al, Nat Commun, 2020)
Architecture of pulmonary lamellar bodies by in cellulo cryo-electron tomography (Klein, Wimmer, et al, Commun Biol, 2021)
Dual-axis Volta phase plate cryo-electron tomography of Ebola virus-like particles (Winter, S.et al, JSB, 2021)
Winter SL, Chlanda P (2021), Dual-axis Volta phase plate cryo-electron tomography of Ebola virus-like particles reveals actin-VP40 interactions, J Struct Biol., doi: 10.1016/j.jsb.2021.107742.
Zhirnov OP, Chlanda P (2021) , The cleavage of spike protein НА0→НА1/HA2 by trypsin permits activation of the M2 channel without its proteolytic cleavage in the influenza A virus, Virology, doi: 10.1016/j.virol.2021.03.016.
Klein S, Wachsmuth-Melm M, Winter SL, Kolovou A, Chlanda P (2021), Cryo-correlative light and electron microscopy workflow for cryo-focused ion beam milled adherent cells, Methods in Cell Biology, doi.org/10.1016/bs.mcb.2020.12.009
Klein S, Wimmer BH, Winter SL, Kolovou A, Laketa V, Chlanda P (2021), Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies. Commun Biol., 4(1):137. doi: 10.1038/s42003-020-01567-z.
Klein S, Cortese M, Winter SL, Wachsmuth-Melm M, Neufeldt CJ, Cerikan B, Stanifer ML, Boulant S, Bartenschlager R, Chlanda P (2020), SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. Nat Commun., 11(1):5885. doi: 10.1038/s41467-020-19619-7.
Klein S, Müller TG, Khalid D, Sonntag-Buck V, Heuser AM, Glass B, Meurer M, Morales I, Schillak A, Freistaedter A, Ambiel I, Winter SL, Zimmermann L, Naumoska T, Bubeck F, Kirrmaier D, Ullrich S, Barreto Miranda I, Anders S, Grimm D, Schnitzler P, Knop M, Kräusslich HG, Dao Thi VL, Börner K, Chlanda P, SARS-CoV-2 RNA Extraction Using Magnetic Beads for Rapid Large-Scale Testing by RT-qPCR and RT-LAMP (2020), Viruses, 12(8):863. doi: 10.3390/v12080863.
Dao Thi VL, Herbst K, Boerner K, Meurer M, Kremer LP, Kirrmaier D, Freistaedter A, Papagiannidis D, Galmozzi C, Stanifer ML, Boulant S, Klein S, Chlanda P, Khalid D, Barreto Miranda I, Schnitzler P, Kräusslich HG, Knop M, Anders S., A colorimetric RT-LAMP assay and LAMP-sequencing for detecting SARS-CoV-2 RNA in clinical samples (2020). Sci Transl Med, 12(556):eabc7075 doi: 10.1126/scitranslmed.abc7075.