PXD022564

PXD022564 is an original dataset announced via ProteomeXchange.

Title	PKR activity modulation by phosphomimetic mutations of serine residues located three aminoacids upstream of double-stranded RNA binding motifs
Description	Eukaryotic translation initiation factor 2 alpha kinase 2 (EIF2AK2), better known as PKR plays a key role in the response to viral infections and cellular homeostasis by regulating mRNA translation. Upon binding dsRNA, PKR is activated through homodimerization and subsequent autophosphorylation on Thr446 and Thr451 residues. In this study, we identified a novel PKR phosphorylation site, Ser6, located 3 aminoacids upstream the first double-stranded RNA binding domain (DRBM1). Another Ser residue occurs in PKR at position 97, the very same position relative to the DRBM2. Ser or Thr residues also occur 3 amino acids upstream DRBMs of other proteins such as ADAR1 or DICER. Phosphoinhibiting mutations (Ser-to-Ala) introduced at Ser6 and Ser97 spontaneously activated PKR. In contrast, phosphomimetic mutations (Ser-to-Asp) inhibited PKR activation following either poly (I:C) transfection or virus infection. These mutations moderately affected dsRNA binding, suggesting a model where negative charges occuring at position 6 and 97 tighten the interaction of DRBMs with the kinase domain, thus keeping PKR in an inactive, closed conformation even in the presence of dsRNA. This study provides new insights on PKR regulation mechanisms and identifies Ser6 and Ser97 as potential targets to modulate PKR activity for therapeutic purposes
HostingRepository	PRIDE
AnnounceDate	2021-05-11
AnnouncementXML	Submission_2021-05-10_22:26:47.936.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD022564
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Didier Vertommen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidoethyl-L-cysteine; phosphorylated residue; N-acetyl-L-methionine; monohydroxylated residue; mono N-acetylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-16 08:01:21	ID requested
1	2021-05-10 00:28:55	announced
2	2021-05-10 22:21:49	announced	2021-05-11: Updated project metadata.
⏵ 3	2021-05-10 22:26:48	announced	2021-05-11: Updated project metadata. 2021-05-11: Updated publication reference for PubMed record(s): 33911136. 2021-05-11: Updated publication reference for DOI(s): 10.1038/S41598-021-88610-Z.

10.1038/S41598-021-88610-Z;

Cesaro T, Hayashi Y, Borghese F, Vertommen D, Wavreil F, Michiels T, PKR activity modulation by phosphomimetic mutations of serine residues located three aminoacids upstream of double-stranded RNA binding motifs. Sci Rep, 11(1):9188(2021) [pubmed]

submitter keyword: dsRNA binding, viral infection,PKR, autophosphorylation, regulation

Thomas Michiels
contact affiliation	Université Catholique de Louvain, de Duve Institute, VIRO B1.74.07, 74, avenue Hippocrate, B-1200, Brussels, Belgium
contact email	thomas.michiels@uclouvain.be
lab head
Didier Vertommen
contact affiliation	UCL - de Duve Institute, Brussels Belgium
contact email	didier.vertommen@uclouvain.be
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/2021/05/PXD022564

PRIDE project URI

• PRIDE
  1. PXD022564
     1. Label: PRIDE project
     2. Name: PKR activity modulation by phosphomimetic mutations of serine residues located three aminoacids upstream of double-stranded RNA binding motifs