PXD045299

PXD045299 is an original dataset announced via ProteomeXchange.

Title	SARS-CoV-2 Envelope TurboID LFQ using internal tagging
Description	SARS-CoV-2 is the causative agent of the Covid-19 pandemic. Here we investigated the cell biology of the SARS-CoV-2 protein Envelope (E). Envelope is a transmembrane protein that oligomerises to form a pentameric cation channel and is essential for high viral titres. One of SARS-CoV-2's four structural proteins (along with Spike (S), Membrane (M), and Nucleocapsid (N)), Envelope is present in the virion membrane. However, most Envelope is not incorporated into the virion, suggesting it has additional functions in disrupting host cell biology. The study by Pearson et al. investigated the trafficking motifs encoded and the host proteins by Envelope to achieve its trafficking itinerary, with the major focus being how Envelope traffics to lysosomes to cause their deacidification. In this study, Pearson et al. investigated the proximal proteome of Envelope by use of TurboID proximity biotinylation proteomics with label-free quantification (LFQ). Envelope was tagged internally which differs from previous approaches which inadvertently disrupt essential trafficking motifs via the position of the affinity or biotinylation tag. The proteomics was performed using Envelope tagged at three different positions (Site3 - 'TAL', Site 4 - 'VNVS', Extreme C-terminus - 'Cterm') and used both wildtype Envelope and Envelope mutants of interest identified through our cell biology studies of key Envelope trafficking motifs ('Delta' - deletion of C-terminal DLLV; DEWV and SVKI - replacement of C-terminal DLLV with indicated amino acids; 'AxA' - R61A and K63A mutations (not included in the final manuscript)). The dataset also includes TurboID alone ('Cyto') as a control. Each condition was assayed in triplicate. These data identified many novel potential interactors of Envelope, some of which have been confirmed by immunoprecipitation biochemical experiments.
HostingRepository	PRIDE
AnnounceDate	2024-02-27
AnnouncementXML	Submission_2024-02-27_07:42:40.689.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Guy Pearson
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-09-12 03:33:35	ID requested
⏵ 1	2024-02-27 07:42:41	announced

Dataset with its publication pending

submitter keyword: SARS-CoV-2, E, TurboID, proximity biotinylation, LFQ, Envelope, BioID

Jeremy Carlton
contact affiliation	Organelle Dynamics Laboratory, The Francis Crick Institute, 1 Midland Road London, NW1 1AT, UK; School of Cancer & Pharmaceutical Sciences, King’s College London, Great Maze Pond, SE1 1UL, UK
contact email	jeremy.carlton@kcl.ac.uk
lab head
Guy Pearson
contact affiliation	Carlton Lab, Kings College London and Francis Crick Institute
contact email	guy.pearson@crick.ac.uk
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/02/PXD045299

PRIDE project URI

• PRIDE
  1. PXD045299
     1. Label: PRIDE project
     2. Name: SARS-CoV-2 Envelope TurboID LFQ using internal tagging