In-situ cryo-electron tomography of Enterovirus replication

a 2023

In-situ cryo-electron tomography of Enterovirus replication

TREBICHALSKÁ, Zuzana, Tibor FÜZIK, Jiří NOVÁČEK a Pavel PLEVKA

Základní údaje

Originální název

In-situ cryo-electron tomography of Enterovirus replication

Autoři

TREBICHALSKÁ, Zuzana, Tibor FÜZIK, Jiří NOVÁČEK a Pavel PLEVKA

Vydání

In situ structural biology: from cryo-EM to multi-scale modelling, 2023

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10607 Virology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

webstranka konference

Organizační jednotka

Středoevropský technologický institut

Příznaky

Mezinárodní význam

Změněno: 6. 3. 2023 12:41, Mgr. Zuzana Trebichalská

Anotace

V originále

Enteroviruses, a large group of non-enveloped picornaviruses, cause a range of human diseases, from the common cold to poliomyelitis. The molecular mechanisms of enterovirus replication and virion assembly are not well understood. Here, we present the cryo-FIB milling and cryo-electron tomography of Echovirus 18-infected cells to characterize the late stages of enterovirus infection in situ. We describe rearrangements of cellular membranes that enable the formation of viral replication factories. Furthermore, we have identified different virus particle assembly intermediates – capsid segments, open and complete capsids, and progeny virions. Some of the virus particles and assembly intermediates were localized in the cytoplasm, whereas others were bound to membranes. We have employed template matching using the structures of the whole virus capsid and a capsid missing three pentamers to identify the complete genome-containing and empty virus particles and virus assembly intermediates, respectively. The particles have been further analysed by subtomogram averaging and classification. Our results provide insight into the virus arrangement during the late stages of the replication cycle of enteroviruses. Additionally, the employed subtomogram averaging pipeline presents a further analysis of viruses and large molecular complexes in situ. We acknowledge the Cryo-electron microscopy and Tomography Core Facility of CEITEC MU for their support in data collection and analysis.

Návaznosti

GX19-25982X, projekt VaV

Název: Analýza replikace enterovirů s využitím elektronové mikroskopie

Investor: Grantová agentura ČR, Structural study of enterovirus replication in situ

MUNI/A/1414/2022, interní kód MU

Název: Specifický výzkum v oblasti „Life Sciences“

Investor: Masarykova univerzita, Specifický výzkum v oblasti „Life Sciences“

Citovat

TREBICHALSKÁ, Zuzana, Tibor FÜZIK, Jiří NOVÁČEK a Pavel PLEVKA. In-situ cryo-electron tomography of Enterovirus replication. In In situ structural biology: from cryo-EM to multi-scale modelling. 2023.

@proceedings{2256457,
   author = {Trebichalská, Zuzana and Füzik, Tibor and Nováček, Jiří and Plevka, Pavel},
   booktitle = {In situ structural biology: from cryo-EM to multi-scale modelling},
   language = {eng},
   title = {In-situ cryo-electron tomography of Enterovirus replication},
   url = {https://www.embl.org/about/info/course-and-conference-office/events/iss23-01/?utm_source=EMBL+courses+and+conferences&utm_campaign=4e2a95e1cf-ISS23-01&utm_medium=email&utm_term=0_b9f46bb4da-4e2a95e1cf-91083209#vf-tabs__section-53de8397-0f08-449c-b4a4-0fd},
   year = {2023}
}

TY  - CONF
ID  - 2256457
AU  - Trebichalská, Zuzana - Füzik, Tibor - Nováček, Jiří - Plevka, Pavel
PY  - 2023
TI  - In-situ cryo-electron tomography of Enterovirus replication
UR  - https://www.embl.org/about/info/course-and-conference-office/events/iss23-01/?utm_source=EMBL+courses+and+conferences&utm_campaign=4e2a95e1cf-ISS23-01&utm_medium=email&utm_term=0_b9f46bb4da-4e2a95e1cf-91083209#vf-tabs__section-53de8397-0f08-449c-b4a4-0fd
N2  - Enteroviruses, a large group of non-enveloped picornaviruses, cause a range of human diseases, from the common cold to poliomyelitis. The molecular mechanisms of enterovirus replication and virion assembly are not well understood. Here, we present the cryo-FIB milling and cryo-electron tomography of Echovirus 18-infected cells to characterize the late stages of enterovirus infection in situ. We describe rearrangements of cellular membranes that enable the formation of viral replication factories. Furthermore, we have identified different virus particle assembly intermediates – capsid segments, open and complete capsids, and progeny virions. Some of the virus particles and assembly intermediates were localized in the cytoplasm, whereas others were bound to membranes. We have employed template matching using the structures of the whole virus capsid and a capsid missing three pentamers to identify the complete genome-containing and empty virus particles and virus assembly intermediates, respectively. The particles have been further analysed by subtomogram averaging and classification. Our results provide insight into the virus arrangement during the late stages of the replication cycle of enteroviruses. Additionally, the employed subtomogram averaging pipeline presents a further analysis of viruses and large molecular complexes in situ. We acknowledge the Cryo-electron microscopy and Tomography Core Facility of CEITEC MU for their support in data collection and analysis.
ER  -

TREBICHALSKÁ, Zuzana, Tibor FÜZIK, Jiří NOVÁČEK a Pavel PLEVKA. In-situ cryo-electron tomography of Enterovirus replication. In \textit{In situ structural biology: from cryo-EM to multi-scale modelling}. 2023.

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