PXD051058-1

PXD051058 is an original dataset announced via ProteomeXchange.

Title	Deciphering the role of the SMG1:SMG8:SMG9 complex during the first authentication step in NMD
Description	Nonsense-mediated mRNA decay (NMD) is a translation-dependent mRNA turnover pathway, which degrades transcripts containing premature termination codons. The execution of NMD requires the phosphorylation of N- and C-terminal tails of the key NMD factor UPF1, which thereby serve as binding platforms for the degradation factors SMG5, SMG6 and SMG7. UPF1 phosphorylation is mediated by the kinase SMG1, whose activity is regulated by a heterodimer consisting of SMG8 and SMG9. Recent work indicated that SMG9 functions as a bridge between SMG1 and SMG8, allowing the C-terminus of SMG8 to elicits its role of stabilizing the autoinhibitory state of SMG1. Here we established SMG8 and SMG9 knockout cell lines in human colorectal adenocarcinoma cell line HCT116 via CRISPR-Cas9. In addition, we used CRISPR-Cas9 to add a FLAG-tag to the N-terminus of endogenous UPF1. With these cell lines we wanted to explore the role of SMG8 and SMG9 for SMG1 regulation and their influence on the UPF1 interactome. Furthermore, we investigated changes in the UPF1 interactome upon treatment with the SMG1 inhibitor SMG1i, which functions as an ATP-competitive inhibitor and binds to the active site of SMG1. Cells were treated with 0, 0.1 or 1 μM SMG1i for 24 h and FLAG co-IP was performed. As controls, the HCT116 wildtype cells were treated with DMSO.
HostingRepository	PRIDE
AnnounceDate	2026-02-13
AnnouncementXML	Submission_2026-02-13_04:11:57.728.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Proteomics Facility
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	monohydroxylated residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-03-28 07:21:04	ID requested
⏵ 1	2026-02-13 04:12:00	announced

Dataset with its publication pending

submitter keyword: SMG8, SMG9, SMG1i, NMD,Nonsense-mediated mRNA decay, UPF1, SMG1

Niels H. Gehring
contact affiliation	Institute for Genetics, University of Cologne, Cologne, Germany
contact email	ngehring@uni-koeln.de
lab head
Proteomics Facility
contact affiliation	CECAD Research Center
contact email	proteomics-facility@uni-koeln.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD051058
     1. Label: PRIDE project
     2. Name: Deciphering the role of the SMG1:SMG8:SMG9 complex during the first authentication step in NMD