PXD054117-1

PXD054117 is an original dataset announced via ProteomeXchange.

Title	SINV-nsP3 mScarelt-tagged immunoprecipitation
Description	1x107 cells HEK293 FLP-IN T-REX cells were infected with SINVnsP3-mScarlet (bait) or SINVwt (negative control) at 1 MOI. After 18 hours, cells were scrapped and spin down at 300 g 4°C for 10 mins. After the centrifugation, supernatants were discarded and cell pellet washed once with 1X PBS, followed by another centrifugation at 300xg 4°C for 10min. After discarding the supernatant, cell pellets were lysed with 200µL ice-cold lysis buffer (10mM Tris-Cl pH 7.5, 150mM NaCl, 0.5mM EDTA, 0.5% IGEPAL, benzonase(1U/mL) and 0.1mM AEBSF serine protease inhibitor) , and incubated on ice for 30min. Then, cell lysates were cleared by centrifugation 17,000xg for 10min at 4°C. The supernatants were transferred to new tubes and diluted 1:1 with the dilution buffer (10mM Tris-Cl pH 7.5 150mM NaCl 0.5mM EDTA 0.05% IGEPAL). Next, cell lysates were incubated with 15µL of RFP-Trap® Magnetic Particles M-270 (pre-washed once with the dilution buffer) for 1 hour at 4 °C with mild rotation. After the incubation, the magnetic beads were washed five times with wash buffer1 (10mM Tris/Cl pH 7.5, 150mM NaCl, 0.5mM EDTA and 0.25% IGEPAL), followed by two washes with the dilution buffer. The proteins elution from the beads was done twice at room temperature with shaking, the first elution with 45µL 0.2M glycine pH 2 solution and the second elution with 45µL 0.2M glycine pH 2 (supplemented with 1%SDS) solution. Finally combined protein eluates were neutralized with 10µL 1M Tris-base pH 10. For proteomic analysis three biological replicates were generated.
HostingRepository	PRIDE
AnnounceDate	2024-12-19
AnnouncementXML	Submission_2024-12-19_14:01:50.420.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wael Kamel
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-07-22 07:57:36	ID requested
⏵ 1	2024-12-19 14:01:51	announced

Kamel W, Ruscica V, Embarc-Buh A, de Laurent ZR, Garcia-Moreno M, Demyanenko Y, Orton RJ, Noerenberg M, Madhusudhan M, Iselin L, J, ä, rvelin AI, Hannan M, Kitano E, Moore S, Merits A, Davis I, Mohammed S, Castello A, Alphavirus infection triggers selective cytoplasmic translocation of nuclear RBPs with moonlighting antiviral roles. Mol Cell, 84(24):4896-4911.e7(2024) [pubmed]

10.1016/j.molcel.2024.11.015;

submitter keyword: SINV-nsP3

Alfredo Castelo
contact affiliation	MRC-University of Glasgow Centre for Virus Research, G61 1QH, Glasgow, Scotland (UK)
contact email	alfredo.castello@glasgow.ac.uk
lab head
Wael Kamel
contact affiliation	Glasgow university
contact email	wael.s.kamel@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/2024/12/PXD054117

PRIDE project URI

• PRIDE
  1. PXD054117
     1. Label: PRIDE project
     2. Name: SINV-nsP3 mScarelt-tagged immunoprecipitation