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1. Robust hepatitis E virus infection and transcriptional response in human hepatocytes

Author

Todt, Daniel, Friesland, Martina, Moeller, Nora, Praditya, Dimas, Kinast, Volker, Brüggemann, Yannick, Knegendorf, Leonard, Burkard, Thomas, Steinmann, Joerg, Burm, Rani, Verhoye, Lieven, Wahid, Avista, Meister, Toni Luise, Engelmann, Michael, Pfankuche, Vanessa M., Puff, Christina, Vondran, Florian W. R., Baumgärtner, Wolfgang, Meuleman, Philip, Behrendt, Patrick, and Steinmann, Eike

Published
2020

2. Landscape of protein-protein interactions during hepatitis C virus assembly and release

Author

Matthaei, Alina, Joecks, Sebastian, Frauenstein, Annika, Bruening, Janina, Bankwitz, Dorothea, Friesland, Martina, Gerold, Gisa, Vieyres, Gabrielle, Kaderali, Lars, Meissner, Felix, Pietschmann, Thomas, Matthaei, Alina, Joecks, Sebastian, Frauenstein, Annika, Bruening, Janina, Bankwitz, Dorothea, Friesland, Martina, Gerold, Gisa, Vieyres, Gabrielle, Kaderali, Lars, Meissner, Felix, and Pietschmann, Thomas

Abstract

Assembly of infectious hepatitis C virus (HCV) particles requires multiple cellular proteins including for instance apolipoprotein E (ApoE). To describe these protein-protein interactions, we performed an affinity purification mass spectrometry screen of HCV-infected cells. We used functional viral constructs with epitope-tagged envelope protein 2 (E2), protein (p) 7, or nonstructural protein 4B (NS4B) as well as cells expressing a tagged variant of ApoE. We also evaluated assembly stage-dependent remodeling of protein complexes by using viral mutants carrying point mutations abrogating particle production at distinct steps of the HCV particle production cascade. Five ApoE binding proteins, 12 p7 binders, 7 primary E2 interactors, and 24 proteins interacting with NS4B were detected. Cell-derived PREB, STT3B, and SPCS2 as well as viral NS2 interacted with both p7 and E2. Only GTF3C3 interacted with E2 and NS4B, highlighting that HCV assembly and replication complexes exhibit largely distinct interactomes. An HCV core protein mutation, preventing core protein decoration of lipid droplets, profoundly altered the E2 interactome. In cells replicating this mutant, E2 interactions with HSPA5, STT3A/B, RAD23A/B, and ZNF860 were significantly enhanced, suggesting that E2 protein interactions partly depend on core protein functions. Bioinformatic and functional studies including STRING network analyses, RNA interference, and ectopic expression support a role of Rad23A and Rad23B in facilitating HCV infectious virus production. Both Rad23A and Rad23B are involved in the endoplasmic reticulum (ER)-associated protein degradation (ERAD). Collectively, our results provide a map of host proteins interacting with HCV assembly proteins, and they give evidence for the involvement of ER protein folding machineries and the ERAD pathway in the late stages of the HCV replication cycle.IMPORTANCEHepatitis C virus (HCV) establishes chronic infections in the majority of exposed individuals.

Published
2024
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5. Virucidal Activity of World Health Organization–Recommended Formulations Against Enveloped Viruses, Including Zika, Ebola, and Emerging Coronaviruses

Author

Siddharta, Anindya, Pfaender, Stephanie, Vielle, Nathalie Jane, Dijkman, Ronald, Friesland, Martina, Becker, Britta, Yang, Jaewon, Engelmann, Michael, Todt, Daniel, Windisch, Marc P., Brill, Florian H., Steinmann, Joerg, Steinmann, Jochen, Becker, Stephan, Alves, Marco P., Pietschmann, Thomas, Eickmann, Markus, Thiel, Volker, and Steinmann, Eike

Published
2017

7. Respiratory Syncytial Virus Two-Step Infection Screen Reveals Inhibitors of Early and Late Life Cycle Stages

Author

Sake, Svenja M., primary, Kosch, Christina, additional, Blockus, Sebastian, additional, Haid, Sibylle, additional, Gunesch, Antonia P., additional, Zhang, Xiaoyu, additional, Friesland, Martina, additional, Trummer, Sofie B., additional, Grethe, Christina, additional, Kühnel, Anne, additional, Rückert, Jessica, additional, Duprex, W. Paul, additional, Huang, Jiabin, additional, Rameix-Welti, Marie-Anne, additional, Empting, Martin, additional, Fischer, Nicole, additional, Hirsch, Anna K. H., additional, Schulz, Thomas F., additional, and Pietschmann, Thomas, additional

Published
2022
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8. Inactivation of Hepatitis C Virus Infectivity by Human Breast Milk

Author

Pfaender, Stephanie, Heyden, Julia, Friesland, Martina, Ciesek, Sandra, Ejaz, Asim, Steinmann, Joerg, Steinmann, Jochen, Malarski, Angelika, Stoiber, Heribert, Tsiavaliaris, Georgios, Bader, Werner, Jahreis, Gerhard, Pietschmann, Thomas, and Steinmann, Eike

Published
2013

9. High tolerance of hepatitis E virus towards alcohol-based disinfectants

Author

Behrendt, Patrick, primary, Friesland, Martina, additional, Wissmann, Jan-Erik, additional, Kinast, Volker, additional, Stahl, Yannick, additional, Praditya, Dimas, additional, Hueffner, Lucas, additional, Nörenberg, Pia Maria, additional, Bremer, Birgit, additional, Maasoumy, Benjamin, additional, Steinmann, Jochen, additional, Becker, Britta, additional, Paulmann, Dajana, additional, Brill, Florian H.H, additional, Steinmann, Joerg, additional, Ulrich, Rainer, additional, Brueggemann, Yannick, additional, Wedemeyer, Heiner, additional, Todt, Daniel, additional, and Steinmann, Eike, additional

Published
2022
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10. TOP-369 HEV replication and genomic diversification in the human kidney

Author

Gömer, André, Wahid, Avista, Hueffner, Lucas, Friesland, Martina, Dinkelborg, Katja, Aliabadi, Elmira, Laue, Fenja, Janshoff, Saskia, Cornberg, Markus, Maasoumy, Benjamin, Pischke, Sven, Müller, Tobias, Bremer, Birgit, Wiesch, Julian Schulze zur, Benckert, Julia, Ulrich, Rainer, Hardtke, Svenja, Dörge, Petra, Vondran, Florian, Lohse, Ansgar, Manns, Michael P., Todt, Daniel, Wedemeyer, Heiner, Pietschmann, Thomas, Steinmann, Eike, and Behrendt, Patrick

Published
2024
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11. Hepatitis E virus is highly resistant to alcohol-based disinfectants

Author

Behrendt, Patrick, primary, Friesland, Martina, additional, Wißmann, Jan-Erik, additional, Kinast, Volker, additional, Stahl, Yannick, additional, Praditya, Dimas, additional, Hueffner, Lucas, additional, Nörenberg, Pia Maria, additional, Bremer, Birgit, additional, Maasoumy, Benjamin, additional, Steinmann, Jochen, additional, Becker, Britta, additional, Paulmann, Dajana, additional, Brill, Florian H.H., additional, Steinmann, Joerg, additional, Ulrich, Rainer G., additional, Brüggemann, Yannick, additional, Wedemeyer, Heiner, additional, Todt, Daniel, additional, and Steinmann, Eike, additional

Published
2022
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12. Viral Interference of Hepatitis C and E Virus Replication in Novel Experimental Co-Infection Systems

Author

Burkard, Thomas, primary, Proske, Nora, additional, Resner, Kathrin, additional, Collignon, Laura, additional, Knegendorf, Leonard, additional, Friesland, Martina, additional, Verhoye, Lieven, additional, Sayed, Ibrahim M., additional, Brüggemann, Yannick, additional, Nocke, Maximilian K., additional, Behrendt, Patrick, additional, Wedemeyer, Heiner, additional, Meuleman, Philip, additional, Todt, Daniel, additional, and Steinmann, Eike, additional

Published
2022
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22. The natural compound silvestrol inhibits hepatitis E virus (HEV) replication in vitro and in vivo

Author

Todt, Daniel, primary, Moeller, Nora, additional, Praditya, Dimas, additional, Kinast, Volker, additional, Friesland, Martina, additional, Engelmann, Michael, additional, Verhoye, Lieven, additional, Sayed, Ibrahim M., additional, Behrendt, Patrick, additional, Dao Thi, Viet Loan, additional, Meuleman, Philip, additional, and Steinmann, Eike, additional

Published
2018
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23. Hepatitis E virus replication and interferon responses in human placental cells

Author

Knegendorf, Leonard, primary, Drave, Svenja A., additional, Dao Thi, Viet Loan, additional, Debing, Yannick, additional, Brown, Richard J. P., additional, Vondran, Florian W. R., additional, Resner, Kathrin, additional, Friesland, Martina, additional, Khera, Tanvi, additional, Engelmann, Michael, additional, Bremer, Birgit, additional, Wedemeyer, Heiner, additional, Behrendt, Patrick, additional, Neyts, Johan, additional, Pietschmann, Thomas, additional, Todt, Daniel, additional, and Steinmann, Eike, additional

Published
2018
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29. Characterization of Hepatitis C virus intra- and intergenotypic chimeras reveals a role of the glycoprotein in virus envelopment

Author

Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Deutsche Forschungsgemeinschaft, Steinmann, Eike, Doerrbecker, Juliane, Friesland, Martina, Riebesehl, Nina, Ginkel, Corinne, Hillung, Julia, Gentzsch, Juliane, Lauber, Chris, Brown, Richard, Frentzen, Anne, Pietschmann, Thomas, Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Deutsche Forschungsgemeinschaft, Steinmann, Eike, Doerrbecker, Juliane, Friesland, Martina, Riebesehl, Nina, Ginkel, Corinne, Hillung, Julia, Gentzsch, Juliane, Lauber, Chris, Brown, Richard, Frentzen, Anne, and Pietschmann, Thomas

Abstract

[EN] Hepatitis C virus (HCV) is highly variable and associated with chronic liver disease. Viral isolates are grouped into seven genotypes (GTs). Accumulating evidence indicates that viral determinants in the core to NS2 proteins modulate the efficiency of virus production. However, the role of the glycoproteins E1 and E2 in this process is currently poorly defined. Therefore, we constructed chimeric viral genomes to explore the role of E1 and E2 in HCV assembly. Comparison of the kinetics and efficiency of particle production by intragenotypic chimeras highlighted core and p7 as crucial determinants for efficient virion release. Glycoprotein sequences, however, had only a minimal impact on this process. In contrast, in the context of intergenotypic HCV chimeras, HCV assembly was profoundly influenced by glycoprotein genes. On the one hand, insertion of GT1a-derived (H77) E1-E2 sequences into a chimeric GT2a virus (Jc1) strongly suppressed virus production. On the other hand, replacement of H77 glycoproteins within the GT1a-GT2a chimeric genome H77/C3 by GT2a-derived (Jc1) E1-E2 increased infectious particle production. Thus, within intergenotypic chimeras, glycoprotein features strongly modulate virus production. Replacement of Jc1 glycoprotein genes by H77-derived E1-E2 did not grossly affect subcellular localization of core, E2, and NS2. However, it caused an accumulation of nonenveloped core protein and increased abundance of nonenveloped core protein structures with slow sedimentation. These findings reveal an important role for the HCV glycoproteins E1 and E2 in membrane envelopment, which likely depends on a genotype-specific interplay with additional viral factors.

Published
2013

30. Characterization of Hepatitis C Virus Intra- and Intergenotypic Chimeras Reveals a Role of the Glycoproteins in Virus Envelopment

Author

Steinmann, Eike, primary, Doerrbecker, Juliane, additional, Friesland, Martina, additional, Riebesehl, Nina, additional, Ginkel, Corinne, additional, Hillung, Julia, additional, Gentzsch, Juliane, additional, Lauber, Chris, additional, Brown, Richard, additional, Frentzen, Anne, additional, and Pietschmann, Thomas, additional

Published
2013
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32. Hepatitis C Virus p7 is Critical for Capsid Assembly and Envelopment

Author

Gentzsch, Juliane, primary, Brohm, Christiane, additional, Steinmann, Eike, additional, Friesland, Martina, additional, Menzel, Nicolas, additional, Vieyres, Gabrielle, additional, Perin, Paula Monteiro, additional, Frentzen, Anne, additional, Kaderali, Lars, additional, and Pietschmann, Thomas, additional

Published
2013
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35. Completion of Hepatitis C Virus Replication Cycle in Heterokaryons Excludes Dominant Restrictions in Human Non-liver and Mouse Liver Cell Lines

Author

Frentzen, Anne, primary, Hüging, Kathrin, additional, Bitzegeio, Julia, additional, Friesland, Martina, additional, Haid, Sibylle, additional, Gentzsch, Juliane, additional, Hoffmann, Markus, additional, Lindemann, Dirk, additional, Zimmer, Gert, additional, Zielecki, Florian, additional, Weber, Friedemann, additional, Steinmann, Eike, additional, and Pietschmann, Thomas, additional

Published
2011
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36. Hepatitis C Virus Hypervariable Region 1 Modulates Receptor Interactions, Conceals the CD81 Binding Site, and Protects Conserved Neutralizing Epitopes

Author

Bankwitz, Dorothea, primary, Steinmann, Eike, additional, Bitzegeio, Julia, additional, Ciesek, Sandra, additional, Friesland, Martina, additional, Herrmann, Eva, additional, Zeisel, Mirjam B., additional, Baumert, Thomas F., additional, Keck, Zhen-yong, additional, Foung, Steven K. H., additional, Pécheur, Eve-Isabelle, additional, and Pietschmann, Thomas, additional

Published
2010
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38. Correspondence.

Author

Steinmann, Eike, Ciesek, Sandra, Friesland, Martina, Erichsen, Thomas J., and Pietschmann, Thomas

Subjects
LETTERS to the editor, HEPATITIS C virus
Abstract

A letter to the editor is presented in response to the article on outbreak of hepatitis C virus infections among people who underwent treatment in an ambulatory specialty clinic, in the August 2010 issue.

Published
2011
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39. C19orf66 is an interferon-induced inhibitor of HCV replication that restricts formation of the viral replication organelle.

Author

Kinast, Volker, Plociennikowska, Agnieszka, Anggakusuma, Bracht, Thilo, Todt, Daniel, Brown, Richard J.P., Boldanova, Tujana, Zhang, Yudi, Brüggemann, Yannick, Friesland, Martina, Engelmann, Michael, Vieyres, Gabrielle, Broering, Ruth, Vondran, Florian W.R., Heim, Markus H., Sitek, Barbara, Bartenschlager, Ralf, Pietschmann, Thomas, and Steinmann, Eike

Subjects
*VIRAL replication, *PROTEOMICS, *CHRONIC hepatitis C, *ZINC-finger proteins, *MASS spectrometry
Abstract

HCV is a positive-strand RNA virus that primarily infects human hepatocytes. Recent studies have reported that C19orf66 is expressed as an interferon (IFN)-stimulated gene; however, the intrinsic regulation of this gene within the liver as well as its antiviral effects against HCV remain elusive. Expression of C19orf66 was quantified in both liver biopsies and primary human hepatocytes, with or without HCV infection. Mechanistic studies of the potent anti-HCV phenotype mediated by C19orf66 were conducted using state-of-the-art virological, biochemical and genetic approaches, as well as correlative light and electron microscopy and transcriptome and proteome analysis. Upregulation of C19orf66 mRNA was observed in both primary human hepatocytes upon HCV infection and in the livers of patients with chronic hepatitis C (CHC). In addition, pegIFNα/ribavirin therapy induced C19orf66 expression in patients with CHC. Transcriptomic profiling and whole cell proteomics of hepatoma cells ectopically expressing C19orf66 revealed no induction of other antiviral genes. Expression of C19orf66 restricted HCV infection, whereas CRIPSPR/Cas9 mediated knockout of C19orf66 attenuated IFN-mediated suppression of HCV replication. Co-immunoprecipitation followed by mass spectrometry identified a stress granule protein-dominated interactome of C19orf66. Studies with subgenomic HCV replicons and an expression system revealed that C19orf66 expression impairs HCV-induced elevation of phosphatidylinositol-4-phosphate, alters the morphology of the viral replication organelle (termed the membranous web) and thereby targets viral RNA replication. C19orf66 is an IFN-stimulated gene, which is upregulated in hepatocytes within the first hours post IFN treatment or HCV infection in vivo. The encoded protein possesses specific antiviral activity against HCV and targets the formation of the membranous web. Our study identifies C19orf66 as an IFN-inducible restriction factor with a novel antiviral mechanism that specifically targets HCV replication. Interferon-stimulated genes are thought to be important to for antiviral immune responses to HCV. Herein, we analysed C19orf66, an interferon-stimulated gene, which appears to inhibit HCV replication. It prevents the HCV-induced elevation of phosphatidylinositol-4-phosphate and alters the morphology of HCV's replication organelle. • C19orf66 is upregulated in vivo upon HCV infection and IFN therapy. • Expression of C19orf66 restricts HCV infection. • C19orf66 alters the morphology of the viral replication organelle. • The zinc finger motif of C19orf66 is crucial for its restriction capacity. [ABSTRACT FROM AUTHOR]

Published
2020
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40. Virucidal activity of 2 alcohol-based formulations proposed as hand rubs by the World Health Organization.

Author

Steinmann J, Becker B, Bischoff B, Paulmann D, Friesland M, Pietschmann T, Steinmann J, and Steinmann E

Subjects
Humans, Time Factors, World Health Organization, Alcohols, Disinfectants, Hand Disinfection methods, Microbial Viability drug effects, Viruses drug effects
Abstract

The virucidal activity of 2 hand rubs proposed by the World Health Organization was studied in a quantitative suspension test for chemical disinfectants and antiseptics in human medicine (EN 14476). These formulations are recommended if no hand rubs with declared microbiological activity are available in health care settings. Formulation I, based on ethanol, inactivated bovine viral diarrhea virus (BVDV), hepatitis C virus (HCV), adenovirus, and murine norovirus as a surrogate for human norovirus. Formulation II, based on isopropyl alcohol, was active only against adenovirus and enveloped viruses, such as BVDV and HCV. Both formulations failed to inactivate poliovirus by 4 log(10) steps within 300 seconds., (Copyright 2010 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.)

Published
2010
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