Mass spectrometry based interaction proteomics has become highly sensitive and quantitative. Using state of the art proteomics, we search for cell surface proteins engaged by human pathogenic viruses. Specifically, we employ label free quantification (LFQ) to define the interactome of known host factors such as the SARS-CoV-2 receptor ACE2 and the promiscuous virus attachment factor TIM-1. In a complementary approach, we use proximity labeling (APEX2) and targeted crosslinking approaches to identify proteins, which interact with viral and host proteins during infection. In follow up experiments, we address the functional role of interaction partners using CRISPR/Cas9 knockout, RNA silencing and blocking techniques.
Our major pathogens of interest are zoonotic, re-emerging, mosquito-borne human pathogens of the alphavirus and phlebovirus genus, including Chikungunya virus and Rift Valley Fever Virus. Identified entry and replication factors will be tested for their specificity to various virus strains and their expression in different human tissues targeted by the virus. Lastly, we will analyze if orthologs in mosquitoes and vertebrates, which serve as transmission and reservoir hosts, also function as host factors. The work will shed light on how zoonotic viruses infect host cells with putative implications for antiviral strategy development.
Margaret Kielian (Albert Einstein College, New York, NY, USA),
Pierre-Yves Lozach (Heidelberg University Hospital, Germany),
Charles M. Rice (Rockefeller University, New York, NY, USA),
Niklas Arnberg (Umea University, Sweden)
DFG (GE 2145/3-2), German Liver Foundation (S163/10135/2017), DAAD, ZIB, Friends of the MHH, Knut and Alice Wallenberg Foundation, Kempe Foundation
Quantitative Proteomics of Uukuniemi Virus-host Cell Interactions Reveals GBF1 as Proviral Host Factor for Phleboviruses.
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Dynamic Dystroglycan Complexes Mediate Cell Entry of Lassa Virus.
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Characterization of the Filovirus-Resistant Cell Line SH-SY5Y Reveals Redundant Role of Cell Surface Entry Factors.
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Fedeli C, Torriani G, Galan-Navarro C, Moraz ML, Moreno H, Gerold G, Kunz S (2018) Axl Can Serve as Entry Factor for Lassa Virus Depending on the Functional Glycosylation of Dystroglycan. J Virol 92(5).
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Gerold G, Bruening J, Pietschmann T. (2015) Decoding protein networks during virus entry by quantitative proteomics. Virus Research. 218:25-39.
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