PXD020339

PXD020339 is an original dataset announced via ProteomeXchange.

Title	Integrative Vectors for Regulated Expression of SARS-CoV-2 Proteins Implicated in RNA Metabolism
Description	Infection with SARS-CoV-2 is expected to result in substantial reorganization of host cell RNA metabolism. We identified 14 proteins that were predicted to interact with host RNAs and/or RNA binding proteins, based on published data for SARS-CoV and SARS-CoV-2. Here, we describe a series of affinity-tagged and codon-optimized expression constructs for each of these 14 proteins. Each viral gene was separately tagged at the N-terminus with Flag-His8, the C-terminus with His8-Flag, or left untagged. The resulting constructs were stably integrated into the HEK293 Flp-In TREx genome. Each viral gene was expressed under the control of an inducible Tet-On promoter, allowing expression levels to be tuned to match physiological conditions during infection. Expression time courses were successfully generated for most of the fusion proteins and quantified by western blot. A few fusion proteins were poorly expressed, whereas others, including Nsp1, Nsp12, and N protein, were toxic unless care was taken to minimize background expression. All plasmids and cells lines are available from Addgene or upon request. We anticipate that availability of these resources will facilitate a more detailed understanding of coronavirus molecular biology.
HostingRepository	PRIDE
AnnounceDate	2020-09-25
AnnouncementXML	Submission_2020-12-23_01:57:27.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Stefan Bresson
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	2020-07-13 17:21:23	ID requested
1	2020-09-25 03:16:42	announced
⏵ 2	2020-12-23 01:57:28	announced	2020-12-23: Updated publication reference for PubMed record(s): 33313418.

Bresson S, Robertson N, Sani E, Turowski TW, Shchepachev V, Kompauerova M, Spanos C, Helwak A, Tollervey D, Integrative vectors for regulated expression of SARS-CoV-2 proteins implicated in RNA metabolism. Wellcome Open Res, 5():261(2020) [pubmed]

ProteomeXchange project tag: Sars-cov-2, Covid-19

submitter keyword: COVID-19

fusion proteins

UV crosslinking

RNA

David Tollervey
contact affiliation	Wellcome Centre for Cell Biology, University of Edinburgh
contact email	d.tollervey@ed.ac.uk
lab head
Stefan Bresson
contact affiliation	University of Edinburgh
contact email	stefan.bresson@gmail.com
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/2020/09/PXD020339

PRIDE project URI

• PRIDE
  1. PXD020339
     1. Label: PRIDE project
     2. Name: Integrative Vectors for Regulated Expression of SARS-CoV-2 Proteins Implicated in RNA Metabolism