Characterization of Phosphorylation- and RNA-Dependent UPF1 Interactors by Quantitative Proteomics

Flury, Valentin; Restuccia, Umberto; Bachi, Angela; Mühlemann, Oliver (2014). Characterization of Phosphorylation- and RNA-Dependent UPF1 Interactors by Quantitative Proteomics. Journal of Proteome Research, 13(6), pp. 3038-3053. 10.1021/pr5002143

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Human up-frameshift 1 (UPF1) is an ATP-dependent RNA helicase and phosphoprotein implicated in several biological processes but is best known for its key function in nonsense-mediated mRNA decay (NMD). Here we employed a combination of stable isotope labeling of amino acids in cell culture experiments to determine by quantitative proteomics UPF1 interactors. We used this approach to distinguish between RNA-mediated and protein-mediated UPF1 interactors and to determine proteins that preferentially bind the hypo- or the hyper-phosphorylated form of UPF1. Confirming and expanding previous studies, we identified the eukaryotic initiation factor 3 (eIF3) as a prominent protein-mediated interactor of UPF1. However, unlike previously reported, eIF3 binds to UPF1 independently of UPF1’s phosphorylation state. Furthermore, our data revealed many nucleus-associated RNA-binding proteins that preferentially associate with hyper-phosphorylated UPF1 in an RNase-sensitive manner, suggesting that UPF1 gets recruited to mRNA and becomes phosphorylated before being exported to the cytoplasm as part of the mRNP.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Mühlemann, Oliver

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1535-3893

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

11 Jun 2014 10:29

Last Modified:

26 Jun 2016 01:52

Publisher DOI:

10.1021/pr5002143

BORIS DOI:

10.7892/boris.52242

URI:

https://boris.unibe.ch/id/eprint/52242

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