TP23: Dynamics of hepatitis B virus morphogenesis and its alterations during chronic infectionIn the serum of patients with chronic hepatitis B one does not only find complete DNA-containing virions, but also virions harbouring “empty” or RNA-containing capsids. The proportion of these forms in chronic carriers varies drastically depending on the intrahepatic viral replication activity and the treatment status. We therefore propose that HBV particles are subject to defined ultrastructural dynamics with a series of thermodynamically driven, temporally and spatially regulated events during particle morphogenesis. This might be key to determining the fate of nucleocapsids in the host cell by deciding between either secretion as enveloped virions or recycling to the nucleus to sustain the pool of viral covalently closed circular DNA, which is the central reservoir for HBV persistence.
Pfister S, Rabl J, Wiegand T, Mattei S, Malär AA, Lecoq L, Seitz S, Bartenschlager R, Böckmann A, Nassal M, Boehringer D, Meier BH. (2023) Structural conservation of HBV-like capsid proteins over hundreds of millions of years despite the shift from non-enveloped to enveloped life-style. Nat Commun. 14:1574. doi.org/10.1038/s41467-023-37068-w.Harati Taji Z, Bielytskyi P, Shein M, Sani MA, Seitz S, Schütz AK. (2022) Transient RNA Interactions Leave a Covalent Imprint on a Viral Capsid Protein. J Am Chem Soc. 18;144(19):8536-8550. doi: 10.1021/jacs.1c12439. Jung S, Jacobs K, Shein M, Schütz AK, Mohr F, Stadler H, Stadler D, Lucko AM, Altstetter SM, Wilsch F, Deng L, Protzer U. (2020). Efficient and reproducible depletion of hepatitis B virus from plasma derived extracellular vesicles. J Extracell Vesicles. 10(2), e12040. https://doi.org/10.1002/jev2.12040Seitz S, Habjanič J, Schütz AK, Bartenschlager R. 2020. The Hepatitis B Virus Envelope Proteins: Molecular Gymnastics Throughout the Viral Life Cycle. 2020. Annu Rev Virol. 2020 Jun 29. doi: 10.1146/annurev-virology-092818-015508 (TP09,TP23)Schütz AK, Hornemann, S, Wälti, MA, Greuter, L, Tiberi, C, Cadalbert, R, Ganter, M, Riek, R, Hammar-ström, P, Nilsson, KPR, Böckmann, A, Aguzzi, AA, Meier, BH. 2018. Binding of polythiophenes to amy-loids: structural mapping of the pharmacophore. ACS Chem Neurosci. 9(3):475-481.Lauber C, Seitz S, Mattei S, Suh A, Beck J, Herstein J, Börold J, Salzburger W, Kaderali L, Briggs J, Bartenschlager R. 2017. Deciphering the origin and evolution of hepatitis B viruses by means of a fami-ly of nonenveloped fish viruses. Cell Host Microbe. 22(3):387-399.e6.Schuetz, AK, Kay, LE 2016. A dynamic molecular basis for malfunction in disease mutants of p97/VCP. eLife. 5:e20143.Seitz S, Iancu C, Volz T, Mier W, Dandri M, Urban S, Bartenschlager R. 2016. A slow maturation pro-cess renders hepatitis B virus infectious. Cell Host Microbe. 20(1):25-35. Schütz, AK, Vagt, T, Huber, M, Ovchinnikova, OY, Cadalbert, R, Wall, J, Güntert, P, Böckmann, A, Glockshuber, R, Meier, BH. 2015. Atomic-resolution three-dimensional structure of amyloid β fibrils bearing the Osaka mutation. Angew Chem Int Ed Engl. 54(1):331-335.Herrmann, US, Schütz, AK, Shirani, H, Huang, D, Saban, D, Nuvolone, M, Li, B, Ballmer, B, Åslund, AK, Mason, JJ, Rushing, E, Budka, H, Nyström, S, Hammarström, P, Böckmann, A, Caflisch, A, Meier, BH, Nilsson, KP, Hornemann, S, Aguzzi, A. 2015. Structure-based drug design identifies polythiophenes as antiprion compounds. Sci Transl Med. 7(299):ra123.Petersen J, Dandri M, Mier W, Lütgehetmann M, Volz T, von Weizsäcker F, Haberkorn U, Fischer L, Pol-lok JM, Erbes B, Seitz S, Urban S. 2008. Prevention of hepatitis B virus infection in vivo by entry inhibi-tors derived from the large envelope protein. Nat Biotechnol. 26(3):335-341.Seitz S, Urban S, Antoni C, Böttcher B. 2007. Cryo-electron microscopy of hepatitis B virions reveals variability in envelope capsid interactions. EMBO J. 26(18):4160-4167.
In the serum of patients chronically infected with hepatitis B virus one does not only find complete DNA-containing virions, but also virions harboring “empty” or RNA-containing capsids. The proportion of these non-canonical forms of virions in chronic carriers varies drastically depending on the intrahepatic viral repli-cation activity and the treatment status. Currently, there is little known about the ultrastructural and biochem-ical foundations of particle morphogenesis giving rise to these non-canonical particles. In earlier research work we identified two morphotypes of hepatitis B virions termed “compact” and “gapped” (Seitz et al., 2007). Compact virions displayed intensive capsid-envelope contacts and an integer bilayer profile of the envelope; in gapped virions the capsid-envelope contacts were lost and the bilayer appeared rearranged. A defined subpopulation of compact particles displayed a unique capsid conformation differing from those known from capsid-like particles expressed in E. coli. Furthermore, circulating virions undergo a distinct maturation process. Virions become released from the host cell in an inactive form and successively devel-op infectivity due to the translocation of the PreS domain of the large envelope protein across the viral membrane (Seitz et al., 2016). Based on these observations, we propose that HBV particles are subject to defined ultrastructural dynamics with a series of thermodynamically driven, temporally and spatially regulat-ed events during particle morphogenesis. This might be key to determining the fate of nucleocapsids in the host cell by deciding between either secretion as enveloped virions or recycling to the nucleus to sustain the pool of viral covalently closed circular DNA, which is the central reservoir for HBV persistence. Moreover, we argue that these ultrastructural dynamics comprise several critical checkpoints pivotal for the varying composition of circulating virions during the course of chronic infection. In the proposed funding period, we will characterize nucleocapsids containing the authentic replication complex by biochemical and structural biology methods (Schütz et al., 2015, 2016, 2018). The nucleocapsids will be arrested in different maturation states and subjected to structure determination by cryo-electron microscopy. We will investigate the impact of any structural changes on the binding competence of nucleocapsids to surface protein segments as prerequisite for capsid envelopment in vitro. Moreover, we will perform thorough biochemical interaction studies to quantitatively describe the binding kinetics of nucleocapsids to the HBV surface glycoproteins. Nuclear magnetic resonance techniques will be applied to resolve the structure of surface protein segments bound to nucleocapsids. The insight gained into structural details and the biochemical parameters of the interactions determining nucleocapsid envelopment will allow us the quantitative characterization and model-ling of the pathways of HBV particle morphogenesis with respect to their variability during chronic infection.
Virus-specific CD8+ T cell responses in hepatitis B virus/ hepatitis D virus co-infection: mechanisms of failure and strategies for restoration
Role of platelets in control and clearance of hepatitis B virus infection via modulation of liver tissue environment and metabolism
Identification of systems properties determining the dynamics of the hepatitis B virus infection cycle and cross-talk to host cell responses
Viral determinants and innate immunity in persistent hepatitis B virus and hepatitis D virus infections
Role of DNA repair, SUMOylation and nuclear bodies in the formation of the hepatitis B virus cccDNA persistence reservoir
Contribution of innate immune responses to persistence versus clearance of hepatitis C virus and hepatitis A virus infections
Combinatorial knock-down/knock-out strategies to reconstitute antiviral im-munity and eliminate persisting hepatitis B virus cccDNA
Impact of interferon-stimulated genes on virus-specific CD8+ T cells and NK cells in viral hepatitis
Impact of the enteric stage of hepatitis A virus and hepatitis E virus on the outcome of liver infection