Dissecting the functions of SMG5, SMG7 and PNRC2 in nonsense-mediated mRNA decay of human cells

Nicholson, Pamela; Gkratsou, Asimina; Josi, Christoph; Colombo, Martino; Mühlemann, Oliver (2018). Dissecting the functions of SMG5, SMG7 and PNRC2 in nonsense-mediated mRNA decay of human cells. RNA - a publication of the RNA Society, 24(4), pp. 557-573. Cold Spring Harbor Laboratory Press 10.1261/rna.063719.117

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The term "nonsense-mediated mRNA decay" (NMD) originally described the degradation of mRNAs with premature translation-termination codons (PTCs), but its meaning has recently been extended to be a translation-dependent post-transcriptional regulator of gene expression affecting 3-10 % of all mRNAs. The degradation of NMD target mRNAs involves both exonucleolytic and endonucleolytic pathways in mammalian cells. While the latter is mediated by the endonuclease SMG6, the former pathway has been reported to require a complex of SMG5-SMG7 or SMG5-PNRC2 binding to UPF1. However, the existence, dominance and mechanistic details of these exonucleolytic pathways are divisive. Therefore, we have investigated the possible exonucleolytic modes of mRNA decay in NMD by examining the roles of UPF1, SMG5, SMG7 and PNRC2 using a combination of functional assays and interaction mapping. Confirming previous work, we detected an interaction between SMG5 and SMG7 and also a functional need for this complex in NMD. In contrast, we found no evidence for the existence of a physical or functional interaction between SMG5 and PNRC2. Instead, we show that UPF1 interacts with PNRC2 and that it triggers 5'-3' exonucleolytic decay of reporter transcripts in tethering assays. PNRC2 interacts mainly with decapping factors and its knockdown does not affect the RNA levels of NMD reporters. We conclude that PNRC2 is probably an important mRNA decapping factor but that it does not appear to be required for NMD.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Nicholson, Pamela; Gkratsou, Asimina; Colombo, Martino and Mühlemann, Oliver

Subjects:

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

ISSN:

1355-8382

Publisher:

Cold Spring Harbor Laboratory Press

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

24 Apr 2018 08:20

Last Modified:

24 Apr 2018 08:30

Publisher DOI:

10.1261/rna.063719.117

BORIS DOI:

10.7892/boris.110130

URI:

https://boris.unibe.ch/id/eprint/110130

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