RT Journal Article
SR Electronic
T1 Coupled protein quality control during nonsense mediated mRNA decay
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.12.22.473893
DO 10.1101/2021.12.22.473893
A1 Inglis, Alison J.
A1 Guna, Alina
A1 Merchán, Ángel Gálvez
A1 Pal, Akshaye
A1 Esantsi, Theodore K.
A1 Keys, Heather R.
A1 Frenkel, Evgeni M.
A1 Oania, Robert
A1 Weissman, Jonathan S.
A1 Voorhees, Rebecca M.
YR 2022
UL http://biorxiv.org/content/early/2022/03/30/2021.12.22.473893.abstract
AB Translation of mRNAs containing premature termination codons (PTCs) can result in truncated protein products with deleterious effects. Nonsense-mediated decay (NMD) is a surveillance path-way responsible for detecting and degrading PTC containing transcripts. While the molecular mechanisms governing mRNA degradation have been extensively studied, the fate of the nascent protein product remains largely uncharacterized. Here, we use a fluorescent reporter system in mammalian cells to reveal a selective degradation pathway specifically targeting the protein product of an NMD mRNA. We show that this process is post-translational, and dependent on an intact ubiquitin proteasome system. To systematically uncover factors involved in NMD-linked protein quality control, we conducted genome-wide flow cytometry-based screens. Our screens recovered known NMD factors, and suggested a lack of dependence on the canonical ribosome-quality control (RQC) pathway. Finally, one of the strongest hits in our screens was the E3 ubiquitin ligase CNOT4, a member of the CCR4-NOT complex, which is involved in initiating mRNA degradation. We show that CNOT4 is involved in NMD coupled protein degradation, and its role depends on a functional RING ubiquitin ligase domain. Our results demonstrate the existence of a targeted pathway for nascent protein degradation from PTC containing mRNAs, and provide a framework for identifying and characterizing factors involved in this process.Competing Interest StatementThe authors have declared no competing interest.