Assembly of infectious Kaposi's sarcoma-associated herpesvirus progeny requires formation of a pORF19 pentamer

Peter Naniima; Eleonora Naimo; Sandra Koch; Ute Curth; Khaled R Alkharsah; Luisa J Ströh; Anne Binz; Jan-Marc Beneke; Benjamin Vollmer; Heike Böning; Eva Maria Borst; Prashant Desai; Jens Bohne; Martin Messerle; Rudolf Bauerfeind; Pierre Legrand; Beate Sodeik; Thomas F Schulz; Thomas Krey

doi:10.1371/journal.pbio.3001423

Assembly of infectious Kaposi's sarcoma-associated herpesvirus progeny requires formation of a pORF19 pentamer

PLoS Biol. 2021 Nov 4;19(11):e3001423. doi: 10.1371/journal.pbio.3001423. eCollection 2021 Nov.

Authors

Peter Naniima^{1

2}, Eleonora Naimo^{1

2}, Sandra Koch^{1

2}, Ute Curth³, Khaled R Alkharsah⁴, Luisa J Ströh¹, Anne Binz^{1

5}, Jan-Marc Beneke⁶, Benjamin Vollmer⁷, Heike Böning¹, Eva Maria Borst¹, Prashant Desai⁸, Jens Bohne¹, Martin Messerle^{1

2

5}, Rudolf Bauerfeind⁹, Pierre Legrand¹⁰, Beate Sodeik^{1

2

5}, Thomas F Schulz^{1

2

5}, Thomas Krey^{1

2

5

6

11}

Affiliations

¹ Institute of Virology, Hannover Medical School, Hannover, Germany.
² German Center for Infection Research (DZIF), partner sites Hannover-Braunschweig and Hamburg-Lübeck-Borstel-Riems, Braunschweig, Germany.
³ Institute for Biophysical Chemistry, Hannover Medical School, Hannover, Germany.
⁴ Department of Microbiology, College of Medicine, Imam Abdulrahman Bin Faisal University (IAU), Dammam, Saudi Arabia.
⁵ Excellence Cluster 2155 RESIST, Hannover Medical School, Hannover, Germany.
⁶ Center of Structural and Cell Biology in Medicine, Institute of Biochemistry, University of Luebeck, Luebeck, Germany.
⁷ Centre for Structural Systems Biology, Leibniz-Institut für Experimentelle Virologie (HPI), Hamburg, Germany.
⁸ Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University, Baltimore, Maryland, United States of America.
⁹ Research Core Unit Laser Microscopy, Hannover Medical School, Hannover, Germany.
¹⁰ Synchrotron SOLEIL, L'Orme des Merisiers, Gif-sur-Yvette, France.
¹¹ Centre for Structural Systems Biology (CSSB), Hamburg, Germany.

Abstract

Herpesviruses cause severe diseases particularly in immunocompromised patients. Both genome packaging and release from the capsid require a unique portal channel occupying one of the 12 capsid vertices. Here, we report the 2.6 Å crystal structure of the pentameric pORF19 of the γ-herpesvirus Kaposi's sarcoma-associated herpesvirus (KSHV) resembling the portal cap that seals this portal channel. We also present the structure of its β-herpesviral ortholog, revealing a striking structural similarity to its α- and γ-herpesviral counterparts despite apparent differences in capsid association. We demonstrate pORF19 pentamer formation in solution and provide insights into how pentamerization is triggered in infected cells. Mutagenesis in its lateral interfaces blocked pORF19 pentamerization and severely affected KSHV capsid assembly and production of infectious progeny. Our results pave the way to better understand the role of pORF19 in capsid assembly and identify a potential novel drug target for the treatment of herpesvirus-induced diseases.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Capsid / chemistry
Conserved Sequence
Crystallography, X-Ray
DNA Packaging
DNA, Viral / genetics
Drosophila
HEK293 Cells
Herpesvirus 8, Human / physiology*
Herpesvirus 8, Human / ultrastructure
Humans
Models, Molecular
Mutagenesis / genetics
Mutant Proteins / metabolism
Open Reading Frames / genetics*
Protein Multimerization*
Viral Proteins / chemistry
Viral Proteins / metabolism*
Virus Assembly / physiology*

Substances

DNA, Viral
Mutant Proteins
Viral Proteins

Grants and funding

This work was supported by funding to TK by the German Center of Infection Research (DZIF, www.dzif.de/en), by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation, www.dfg.de) under Germany’s Excellence Strategy – EXC 2155 – Projektnummer 390874280, by the Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 158989968 - SFB 900, project B10 and by funding to EMB and TK by the Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 441233738. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.