---
_id: '10639'
abstract:
- lang: eng
text: With more than 80 members worldwide, the Orthobunyavirus genus in the Peribunyaviridae
family is a large genus of enveloped RNA viruses, many of which are emerging pathogens
in humans and livestock. How orthobunyaviruses (OBVs) penetrate and infect mammalian
host cells remains poorly characterized. Here, we investigated the entry mechanisms
of the OBV Germiston (GERV). Viral particles were visualized by cryo-electron
microscopy and appeared roughly spherical with an average diameter of 98 nm. Labeling
of the virus with fluorescent dyes did not adversely affect its infectivity and
allowed the monitoring of single particles in fixed and live cells. Using this
approach, we found that endocytic internalization of bound viruses was asynchronous
and occurred within 30-40 min. The virus entered Rab5a+ early endosomes and, subsequently,
late endosomal vacuoles containing Rab7a but not LAMP-1. Infectious entry did
not require proteolytic cleavage, and endosomal acidification was sufficient and
necessary for viral fusion. Acid-activated penetration began 15-25 min after initiation
of virus internalization and relied on maturation of early endosomes to late endosomes.
The optimal pH for viral membrane fusion was slightly below 6.0, and penetration
was hampered when the potassium influx was abolished. Overall, our study provides
real-time visualization of GERV entry into host cells and demonstrates the importance
of late endosomal maturation in facilitating OBV penetration.
acknowledged_ssus:
- _id: EM-Fac
acknowledgement: This work was supported by INRAE starter funds, Project IDEXLYON (University of Lyon)
within the Programme Investissements d’Avenir (ANR-16-IDEX-0005), and FINOVIAO14
(Fondation pour l’Université de Lyon), all to P.Y.L. This work was also supported by
CellNetworks Research Group funds and Deutsche Forschungsgemeinschaft (DFG) funding
(grant numbers LO-2338/1-1 and LO-2338/3-1) awarded to P.Y.L., Austrian Science Fund
(FWF) grant P31445 to F.K.M.S., a Chinese Scholarship Council (CSC;no. 201904910701)
fellowship to Q.X., and a ministére de l’enseignement supérieur, de la recherche et de
l’innovation (MESRI) doctoral thesis grant to M.D.
article_number: e02146-21
article_processing_charge: No
article_type: original
author:
- first_name: Stefan
full_name: Windhaber, Stefan
last_name: Windhaber
- first_name: Qilin
full_name: Xin, Qilin
last_name: Xin
- first_name: Zina M.
full_name: Uckeley, Zina M.
last_name: Uckeley
- first_name: Jana
full_name: Koch, Jana
last_name: Koch
- first_name: Martin
full_name: Obr, Martin
id: 4741CA5A-F248-11E8-B48F-1D18A9856A87
last_name: Obr
orcid: 0000-0003-1756-6564
- first_name: Céline
full_name: Garnier, Céline
last_name: Garnier
- first_name: Catherine
full_name: Luengo-Guyonnot, Catherine
last_name: Luengo-Guyonnot
- first_name: Maëva
full_name: Duboeuf, Maëva
last_name: Duboeuf
- first_name: Florian KM
full_name: Schur, Florian KM
id: 48AD8942-F248-11E8-B48F-1D18A9856A87
last_name: Schur
orcid: 0000-0003-4790-8078
- first_name: Pierre-Yves
full_name: Lozach, Pierre-Yves
last_name: Lozach
citation:
ama: Windhaber S, Xin Q, Uckeley ZM, et al. The Orthobunyavirus Germiston enters
host cells from late endosomes. Journal of Virology. 2022;96(5). doi:10.1128/jvi.02146-21
apa: Windhaber, S., Xin, Q., Uckeley, Z. M., Koch, J., Obr, M., Garnier, C., … Lozach,
P.-Y. (2022). The Orthobunyavirus Germiston enters host cells from late endosomes.
Journal of Virology. American Society for Microbiology. https://doi.org/10.1128/jvi.02146-21
chicago: Windhaber, Stefan, Qilin Xin, Zina M. Uckeley, Jana Koch, Martin Obr, Céline
Garnier, Catherine Luengo-Guyonnot, Maëva Duboeuf, Florian KM Schur, and Pierre-Yves
Lozach. “The Orthobunyavirus Germiston Enters Host Cells from Late Endosomes.”
Journal of Virology. American Society for Microbiology, 2022. https://doi.org/10.1128/jvi.02146-21.
ieee: S. Windhaber et al., “The Orthobunyavirus Germiston enters host cells
from late endosomes,” Journal of Virology, vol. 96, no. 5. American Society
for Microbiology, 2022.
ista: Windhaber S, Xin Q, Uckeley ZM, Koch J, Obr M, Garnier C, Luengo-Guyonnot
C, Duboeuf M, Schur FK, Lozach P-Y. 2022. The Orthobunyavirus Germiston enters
host cells from late endosomes. Journal of Virology. 96(5), e02146-21.
mla: Windhaber, Stefan, et al. “The Orthobunyavirus Germiston Enters Host Cells
from Late Endosomes.” Journal of Virology, vol. 96, no. 5, e02146-21, American
Society for Microbiology, 2022, doi:10.1128/jvi.02146-21.
short: S. Windhaber, Q. Xin, Z.M. Uckeley, J. Koch, M. Obr, C. Garnier, C. Luengo-Guyonnot,
M. Duboeuf, F.K. Schur, P.-Y. Lozach, Journal of Virology 96 (2022).
date_created: 2022-01-18T10:04:18Z
date_published: 2022-03-01T00:00:00Z
date_updated: 2026-06-18T08:44:25Z
day: '01'
ddc:
- '570'
department:
- _id: FlSc
doi: 10.1128/jvi.02146-21
external_id:
isi:
- '000779305000033'
pmid:
- '35019710'
fulldoi: https://doi.org/10.1128/jvi.02146-21
intvolume: ' 96'
isi: 1
issue: '5'
keyword:
- virology
- insect science
- immunology
- microbiology
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8906410
month: '03'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 26736D6A-B435-11E9-9278-68D0E5697425
call_identifier: FWF
grant_number: P31445
name: Structural conservation and diversity in retroviral capsid
publication: Journal of Virology
publication_identifier:
eissn:
- 1098-5514
issn:
- 0022-538X
publication_status: published
publisher: American Society for Microbiology
quality_controlled: '1'
scopus_import: '1'
status: public
title: The Orthobunyavirus Germiston enters host cells from late endosomes
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 96
year: '2022'
...
---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
_id: '813'
abstract:
- lang: eng
text: The Gag polyprotein of retroviruses drives immature virus assembly by forming
hexameric protein lattices. The assembly is primarily mediated by protein-protein
interactions between capsid (CA) domains and by interactions between nucleocapsid
(NC) domains and RNA. Specific interactions between NC and the viral RNA are required
for genome packaging. Previously reported cryoelectron microscopy analysis of
immature Mason-Pfizer monkey virus (M-PMV) particles suggested that a basic region
(residues RKK) in CA may serve as an additional binding site for nucleic acids.
Here, we have introduced mutations into the RKK region in both bacterial and proviral
M-PMV vectors and have assessed their impact on M-PMV assembly, structure, RNA
binding, budding/release, nuclear trafficking, and infectivity using in vitro
and in vivo systems. Our data indicate that the RKK region binds and structures
nucleic acid that serves to promote virus particle assembly in the cytoplasm.
Moreover, the RKK region appears to be important for recruitment of viral genomic
RNA into Gag particles, and this function could be linked to changes in nuclear
trafficking. Together these observations suggest that in M-PMV, direct interactions
between CA and nucleic acid play important functions in the late stages of the
viral life cycle.
acknowledgement: "Work in the laboratory of John A. G. Briggs was funded by Deutsche\r\nForschungsgemeinschaft
(DFG) (BR 3635/2-1). This work, including the\r\nefforts of Tomas Ruml, was funded
by the Grant Agency of the Czech\r\nRepublic (14-15326S) and the Czech Ministry
of Education (NPU I sus-\r\ntainability projects LO1302 and LO1304)."
article_processing_charge: No
article_type: original
author:
- first_name: Tibor
full_name: Füzik, Tibor
last_name: Füzik
- first_name: Růžena
full_name: Píchalová, Růžena
last_name: Píchalová
- first_name: Florian
full_name: Schur, Florian
id: 48AD8942-F248-11E8-B48F-1D18A9856A87
last_name: Schur
orcid: 0000-0003-4790-8078
- first_name: Karolína
full_name: Strohalmová, Karolína
last_name: Strohalmová
- first_name: Ivana
full_name: Křížová, Ivana
last_name: Křížová
- first_name: Romana
full_name: Hadravová, Romana
last_name: Hadravová
- first_name: Michaela
full_name: Rumlová, Michaela
last_name: Rumlová
- first_name: John
full_name: Briggs, John
last_name: Briggs
- first_name: Pavel
full_name: Ulbrich, Pavel
last_name: Ulbrich
- first_name: Tomáš
full_name: Ruml, Tomáš
last_name: Ruml
citation:
ama: Füzik T, Píchalová R, Schur FK, et al. Nucleic acid binding by Mason-Pfizer
monkey virus CA promotes virus assembly and genome packaging. Journal of Virology.
2016;90(9):4593-4603. doi:10.1128/JVI.03197-15
apa: Füzik, T., Píchalová, R., Schur, F. K., Strohalmová, K., Křížová, I., Hadravová,
R., … Ruml, T. (2016). Nucleic acid binding by Mason-Pfizer monkey virus CA promotes
virus assembly and genome packaging. Journal of Virology. American Society
for Microbiology. https://doi.org/10.1128/JVI.03197-15
chicago: Füzik, Tibor, Růžena Píchalová, Florian KM Schur, Karolína Strohalmová,
Ivana Křížová, Romana Hadravová, Michaela Rumlová, John Briggs, Pavel Ulbrich,
and Tomáš Ruml. “Nucleic Acid Binding by Mason-Pfizer Monkey Virus CA Promotes
Virus Assembly and Genome Packaging.” Journal of Virology. American Society
for Microbiology, 2016. https://doi.org/10.1128/JVI.03197-15.
ieee: T. Füzik et al., “Nucleic acid binding by Mason-Pfizer monkey virus
CA promotes virus assembly and genome packaging,” Journal of Virology,
vol. 90, no. 9. American Society for Microbiology, pp. 4593–4603, 2016.
ista: Füzik T, Píchalová R, Schur FK, Strohalmová K, Křížová I, Hadravová R, Rumlová
M, Briggs J, Ulbrich P, Ruml T. 2016. Nucleic acid binding by Mason-Pfizer monkey
virus CA promotes virus assembly and genome packaging. Journal of Virology. 90(9),
4593–4603.
mla: Füzik, Tibor, et al. “Nucleic Acid Binding by Mason-Pfizer Monkey Virus CA
Promotes Virus Assembly and Genome Packaging.” Journal of Virology, vol.
90, no. 9, American Society for Microbiology, 2016, pp. 4593–603, doi:10.1128/JVI.03197-15.
short: T. Füzik, R. Píchalová, F.K. Schur, K. Strohalmová, I. Křížová, R. Hadravová,
M. Rumlová, J. Briggs, P. Ulbrich, T. Ruml, Journal of Virology 90 (2016) 4593–4603.
date_created: 2018-12-11T11:48:38Z
date_published: 2016-05-01T00:00:00Z
date_updated: 2026-05-20T07:34:00Z
day: '01'
doi: 10.1128/JVI.03197-15
extern: '1'
external_id:
pmid:
- '26912613'
fulldoi: https://doi.org/10.1128/JVI.03197-15
intvolume: ' 90'
issue: '9'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.1128/jvi.03197-15
month: '05'
oa: 1
oa_version: Published Version
page: 4593 - 4603
pmid: 1
publication: Journal of Virology
publication_identifier:
eissn:
- 1098-5514
issn:
- 0022-538X
publication_status: published
publisher: American Society for Microbiology
publist_id: '6835'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Nucleic acid binding by Mason-Pfizer monkey virus CA promotes virus assembly
and genome packaging
type: journal_article
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
volume: 90
year: '2016'
...
---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
_id: '815'
abstract:
- lang: eng
text: "The polyprotein Gag is the primary structural component of retroviruses.
Gag consists of independently folded domains connected by flexible linkers. Interactions
between the conserved capsid (CA) domains of Gag mediate formation of hexameric
protein lattices that drive assembly of immature virus particles. Proteolytic
cleavage of Gag by the viral protease (PR) is required for maturation of retroviruses
from an immature form into an infectious form. Within the assembled Gag lattices
of HIV-1 and Mason- Pfizer monkey virus (M-PMV), the C-terminal domain of CA adopts
similar quaternary arrangements, while the N-terminal domain of CA is packed in
very different manners. Here, we have used cryo-electron tomography and subtomogram
averaging to study in vitro-assembled, immature virus-like Rous sarcoma virus
(RSV) Gag particles and have determined the structure of CA and the surrounding
regions to a resolution of ~8 Å. We found that the C-terminal domain of RSV CA
is arranged similarly to HIV-1 and M-PMV, whereas the N-terminal domain of CA
adopts a novel arrangement in which the upstream p10 domain folds back into the
CA lattice. In this position the cleavage site between CA and p10 appears to be
inaccessible to PR. Below CA, an extended density is consistent with the presence
of a six-helix bundle formed by the spacer-peptide region. We have also assessed
the affect of lattice assembly on proteolytic processing by exogenous PR. The
cleavage between p10 and CA is indeed inhibited in the assembled lattice, a finding
consistent with structural regulation of proteolytic maturation.\r\n"
article_processing_charge: No
article_type: original
author:
- first_name: Florian
full_name: Schur, Florian
id: 48AD8942-F248-11E8-B48F-1D18A9856A87
last_name: Schur
orcid: 0000-0003-4790-8078
- first_name: Robert
full_name: Dick, Robert
last_name: Dick
- first_name: Wim
full_name: Hagen, Wim
last_name: Hagen
- first_name: Volker
full_name: Vogt, Volker
last_name: Vogt
- first_name: John
full_name: Briggs, John
last_name: Briggs
citation:
ama: Schur FK, Dick R, Hagen W, Vogt V, Briggs J. The structure of immature virus
like Rous sarcoma virus gag particles reveals a structural role for the p10 domain
in assembly. Journal of Virology. 2015;89(20):10294-10302. doi:10.1128/JVI.01502-15
apa: Schur, F. K., Dick, R., Hagen, W., Vogt, V., & Briggs, J. (2015). The structure
of immature virus like Rous sarcoma virus gag particles reveals a structural role
for the p10 domain in assembly. Journal of Virology. ASM. https://doi.org/10.1128/JVI.01502-15
chicago: Schur, Florian KM, Robert Dick, Wim Hagen, Volker Vogt, and John Briggs.
“The Structure of Immature Virus like Rous Sarcoma Virus Gag Particles Reveals
a Structural Role for the P10 Domain in Assembly.” Journal of Virology.
ASM, 2015. https://doi.org/10.1128/JVI.01502-15.
ieee: F. K. Schur, R. Dick, W. Hagen, V. Vogt, and J. Briggs, “The structure of
immature virus like Rous sarcoma virus gag particles reveals a structural role
for the p10 domain in assembly,” Journal of Virology, vol. 89, no. 20.
ASM, pp. 10294–10302, 2015.
ista: Schur FK, Dick R, Hagen W, Vogt V, Briggs J. 2015. The structure of immature
virus like Rous sarcoma virus gag particles reveals a structural role for the
p10 domain in assembly. Journal of Virology. 89(20), 10294–10302.
mla: Schur, Florian KM, et al. “The Structure of Immature Virus like Rous Sarcoma
Virus Gag Particles Reveals a Structural Role for the P10 Domain in Assembly.”
Journal of Virology, vol. 89, no. 20, ASM, 2015, pp. 10294–302, doi:10.1128/JVI.01502-15.
short: F.K. Schur, R. Dick, W. Hagen, V. Vogt, J. Briggs, Journal of Virology 89
(2015) 10294–10302.
date_created: 2018-12-11T11:48:39Z
date_published: 2015-09-22T00:00:00Z
date_updated: 2026-05-18T12:47:51Z
day: '22'
doi: 10.1128/JVI.01502-15
extern: '1'
external_id:
pmid:
- '26223638'
fulldoi: https://doi.org/10.1128/JVI.01502-15
intvolume: ' 89'
issue: '20'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.1128/jvi.01502-15
month: '09'
oa: 1
oa_version: Published Version
page: 10294 - 10302
pmid: 1
publication: Journal of Virology
publication_identifier:
eissn:
- 1098-5514
issn:
- 0022-538X
publication_status: published
publisher: ASM
publist_id: '6837'
quality_controlled: '1'
scopus_import: '1'
status: public
title: The structure of immature virus like Rous sarcoma virus gag particles reveals
a structural role for the p10 domain in assembly
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: ba8df636-2132-11f1-aed0-ed93e2281fdd
volume: 89
year: '2015'
...