A Conserved Cysteine Residue in Coxsackievirus B3 Protein 3A with Implication for Elevated Virulence
Martin Voss,
Sandra Pinkert,
Meike Kespohl,
Niclas Gimber,
Karin Klingel,
Jan Schmoranzer,
Michael Laue,
Matthias Gaida,
Peter-Michael Kloetzel,
Antje Beling
Affiliations
Martin Voss
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Sandra Pinkert
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Meike Kespohl
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Niclas Gimber
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Karin Klingel
Cardiopathology, Institute for Pathology and Neuropathology, University of Tübingen, 72016 Tübingen, Germany
Jan Schmoranzer
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Michael Laue
Robert Koch Institute, Advanced Light and Electron Microscopy (ZBS 4), 13353 Berlin, Germany
Matthias Gaida
Institute of Pathology, University Medical Center Mainz, JGU-Mainz, 55131 Mainz, Germany
Peter-Michael Kloetzel
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Antje Beling
Institute of Biochemistry, Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 10117 Berlin, Germany
Enteroviruses (EV) are implicated in an extensive range of clinical manifestations, such as pancreatic failure, cardiovascular disease, hepatitis, and meningoencephalitis. We recently reported on the biochemical properties of the highly conserved cysteine residue at position 38 (C38) of enteroviral protein 3A and demonstrated a C38-mediated homodimerization of the Coxsackievirus B3 protein 3A (CVB3-3A) that resulted in its profound stabilization. Here, we show that residue C38 of protein 3A supports the replication of CVB3, a clinically relevant member of the enterovirus genus. The infection of HeLa cells with protein 3A cysteine 38 to alanine mutants (C38A) attenuates virus replication, resulting in comparably lower virus particle formation. Consistently, in a mouse infection model, the enhanced virus propagation of CVB3-3A wt in comparison to the CVB3-3A[C38A] mutant was confirmed and found to promote severe liver tissue damage. In contrast, infection with the CVB3-3A[C38A] mutant mitigated hepatic tissue injury and ameliorated the signs of systemic inflammatory responses, such as hypoglycemia and hypothermia. Based on these data and our previous report on the C38-mediated stabilization of the CVB3-3A protein, we conclude that the highly conserved amino acid C38 in protein 3A enhances the virulence of CVB3.
