---
OA_place: publisher
_id: '18766'
abstract:
- lang: eng
  text: Poxviruses are large pleomorphic double-stranded DNA viruses that include
    well known members such as variola virus, the causative agent of smallpox, Mpox
    virus, as well as Vaccinia virus (VACV), which serves as a vaccination strain
    for formerly mentioned viruses. VACV is a valuable model for studying large pleomorphic
    DNA viruses in general and poxviruses specifically, as many features, such as
    core morphology and structural proteins, are well conserved within this family.
    Despite decades of research, our understanding of the structural components and
    proteins that comprise the poxvirus core in mature virions remains limited. Although
    major core proteins were identified via indirect experimental evidence, the core's
    complexity, with its large size, structure and number of involved proteins, has
    hindered efforts to achieve high-resolution insights and to define the roles of
    the individual proteins. The specific protein composition of the core's individual
    layers, including the palisade layer and the inner core wall, has remained unclear.
    In this study, we have merged multiple approaches, including single particle cryo
    electron microscopy of purified virus cores, cryo-electron tomography and subtomogram
    averaging of mature virions and molecular modeling to elucidate the structural
    determinants of the VACV core. Due to the lack of experimentally derived structures,
    either in situ or reconstituted in vitro, we used Alphafold to predict models
    of the putative major core protein candidates, A10, 23k, A3, A4, and L4. Our results
    show that the VACV core is composed of several layers with varying local symmetries,
    forming more intricate interactions than observed previously. This allowed us
    to identify several molecular building blocks forming the viral core lattice.
    In particular, we identified trimers of protein A10 as a major core structure
    that forms the palisade layer of the viral core. Additionally, we revealed that
    six petals of a flower shaped core pore within the core wall are composed of A10
    trimers. Furthermore, we obtained a cryo-EM density for the inner core wall that
    could potentially accommodate an A3 dimer. Integrating descriptions of protein
    interactions from previous studies enabled us to provide a detailed structural
    model of the poxvirus core wall, and our findings indicate that the interactions
    within A10 trimers are likely consistent across orthopox- and parapoxviruses.
    This combined application of cryo-SPA and cryo-ET can help overcome obstacles
    in studying complex virus structures in the future, including their key assembly
    proteins, interactions, and the formation into a core lattice. Our work provides
    important fundamental new insights into poxvirus core architecture, also considering
    the recent re-emergence of poxviruses.
acknowledged_ssus:
- _id: EM-Fac
- _id: LifeSc
- _id: ScienComp
acknowledgement: "This work was funded by the Austrian Science Fund (FWF) grant P31445
  and ISTA. I\r\nwould like to express my gratitude to the Scientific Service Units,
  particularly the Lab\r\nSupport Facility, the Scientific Computing Facility and
  the Electron Microscopy Facility\r\nfor their tremendous support. I want to especially
  thank Alois for assisting me with the\r\ninstallation of countless new software
  and for troubleshooting cluster issues. A special\r\nthanks goes to Valentin for
  his outstanding support in cryo-EM data acquisition and\r\nhis ongoing help in improving
  the process to ensure that I obtained the best possible\r\ndata from my sample."
alternative_title:
- ISTA thesis
article_processing_charge: No
author:
- first_name: Julia
  full_name: Datler, Julia
  id: 3B12E2E6-F248-11E8-B48F-1D18A9856A87
  last_name: Datler
  orcid: 0000-0002-3616-8580
citation:
  ama: Datler J. Elucidating the structural determinants of the poxvirus core using
    multi-modal cryo-EM. 2024. doi:<a href="https://doi.org/10.15479/at:ista:18766">10.15479/at:ista:18766</a>
  apa: Datler, J. (2024). <i>Elucidating the structural determinants of the poxvirus
    core using multi-modal cryo-EM</i>. Institute of Science and Technology Austria.
    <a href="https://doi.org/10.15479/at:ista:18766">https://doi.org/10.15479/at:ista:18766</a>
  chicago: Datler, Julia. “Elucidating the Structural Determinants of the Poxvirus
    Core Using Multi-Modal Cryo-EM.” Institute of Science and Technology Austria,
    2024. <a href="https://doi.org/10.15479/at:ista:18766">https://doi.org/10.15479/at:ista:18766</a>.
  ieee: J. Datler, “Elucidating the structural determinants of the poxvirus core using
    multi-modal cryo-EM,” Institute of Science and Technology Austria, 2024.
  ista: Datler J. 2024. Elucidating the structural determinants of the poxvirus core
    using multi-modal cryo-EM. Institute of Science and Technology Austria.
  mla: Datler, Julia. <i>Elucidating the Structural Determinants of the Poxvirus Core
    Using Multi-Modal Cryo-EM</i>. Institute of Science and Technology Austria, 2024,
    doi:<a href="https://doi.org/10.15479/at:ista:18766">10.15479/at:ista:18766</a>.
  short: J. Datler, Elucidating the Structural Determinants of the Poxvirus Core Using
    Multi-Modal Cryo-EM, Institute of Science and Technology Austria, 2024.
corr_author: '1'
date_created: 2025-01-07T10:23:12Z
date_published: 2024-12-30T00:00:00Z
date_updated: 2026-04-07T12:59:44Z
day: '30'
ddc:
- '570'
degree_awarded: PhD
department:
- _id: GradSch
- _id: FlSc
doi: 10.15479/at:ista:18766
file:
- access_level: closed
  checksum: 3e51cab327c754045c3d29c1a50cc9a9
  content_type: application/vnd.openxmlformats-officedocument.wordprocessingml.document
  creator: jstanger
  date_created: 2025-01-07T12:15:11Z
  date_updated: 2025-01-07T12:15:11Z
  file_id: '18769'
  file_name: PhD_thesis_Julia_Datler.docx
  file_size: 38814932
  relation: source_file
- access_level: open_access
  checksum: 22fabe5b97950bf852212f6edb555173
  content_type: application/pdf
  creator: jstanger
  date_created: 2025-01-07T12:15:14Z
  date_updated: 2025-01-07T12:15:14Z
  file_id: '18770'
  file_name: PhD_thesis_Julia_Datler.pdf
  file_size: 12044865
  relation: main_file
  success: 1
file_date_updated: 2025-01-07T12:15:14Z
has_accepted_license: '1'
keyword:
- cryo-EM
- cryo-ET
- cryo-SPA
- Structural Virology
- Poxvirus
- Vaccinia Virus
- Structural Biology
language:
- iso: eng
license: https://creativecommons.org/licenses/by-nc-nd/4.0/
month: '12'
oa: 1
oa_version: Published Version
page: '106'
project:
- _id: 26736D6A-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: P31445
  name: Structural conservation and diversity in retroviral capsid
publication_identifier:
  isbn:
  - 978-3-99078-049-7
  issn:
  - 2663-337X
publication_status: published
publisher: Institute of Science and Technology Austria
related_material:
  record:
  - id: '12334'
    relation: part_of_dissertation
    status: public
  - id: '14979'
    relation: part_of_dissertation
    status: public
status: public
supervisor:
- first_name: Florian KM
  full_name: Schur, Florian KM
  id: 48AD8942-F248-11E8-B48F-1D18A9856A87
  last_name: Schur
  orcid: 0000-0003-4790-8078
title: Elucidating the structural determinants of the poxvirus core using multi-modal
  cryo-EM
tmp:
  image: /images/cc_by_nc_nd.png
  legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
  name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
    (CC BY-NC-ND 4.0)
  short: CC BY-NC-ND (4.0)
type: dissertation
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
year: '2024'
...