Proteomics

Dataset Information

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Proximity biotinylation and LC-MSMS identification of the molecular microenvironment of coronavirus replication complexes

ABSTRACT: To investigate virus-host interactions at the site of coronavirus replication, we developed a biotin-based proximity labelling approach by engineering a promiscuous biotin ligase, BirA-R118G, within the coronavirus replication and transcription complex (RTC). BirA-R118G was fused to non-structural protein 2 to generate MHV- BirA-R118G-nsp2. During the course of infection, viral and host factors in proximity of the viral RTC become covalently biotinylated, allowing affinity purification of biotin-tagged factors and mass spectrometry identification of RTC-proximal viral and host factors. Results provide a comprehensive library of RTC-associated factors which likely include crucial factors that promote, orchestrate and assist viral replication within the viral RTC microenvironment.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Murine Hepatitis Virus Strain A59 Mus Musculus (mouse)

TISSUE(S): Fibroblast

SUBMITTER: Manfred Heller  

LAB HEAD: Volker Thiel

PROVIDER: PXD009975 | Pride | 2019-01-16

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
20161123_PV.zip Other
BiotinKSites.txt Txt
MurineHepatitisVirus.fasta Fasta
MusMusculus_2016_04.fasta Fasta
OxidationMSites.txt Txt
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Publications

Positive-sense RNA viruses hijack intracellular membranes that provide niches for viral RNA synthesis and a platform for interactions with host proteins. However, little is known about host factors at the interface between replicase complexes and the host cytoplasm. We engineered a biotin ligase into a coronaviral replication/transcription complex (RTC) and identified >500 host proteins constituting the RTC microenvironment. siRNA-silencing of each RTC-proximal host factor demonstrated importanc  ...[more]

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