Functional analysis of the grapevine paralogs of the SMG7 NMD factor using a heterolog VIGS-based gene depletion-complementation system
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Nonsense-mediated mRNA decay (NMD) is a eukaryotic quality control system that identifies and eliminates transcripts having a premature translation termination codon (PTC). NMD is also involved in the control of several wild-type mRNAs. The NMD core machinery consists of three highly conserved NMD factors (UPF1, UPF2 and UPF3) and at least one less conserved 14-3-3-like domain containing protein (SMG7). A PTC is identified by UPF factors, and then SMG7 triggers rapid transcript decay. UPF factors are generally encoded by a single gene, whereas SMG7 has duplicated several times during evolution. Recently it was reported that the plant SMG7 is autoregulated through NMD and that SMG7 has two relatively divergent paralogs in dicots, SMG7 and SMG7L. In mammals all three SMG7 related genes (SMG5, SMG6 and SMG7) are essential in NMD, so we hypothesized that in plants the SMG7 and SMG7L duplicates may also play distinct roles in NMD. To test this possibility, we have analyzed the evolution and the function of plant SMG7 homologs. We show that SMG7L is not required for plant NMD. Interestingly, we found that the grapevine and poplar genomes contain two quite divergent SMG7 paralogs which may have derived from an ancient duplication event. Using heterolog depletion/complementation assays we demonstrate that both grapevine SMG7 copies retained the complete NMD activity and both of them are under NMD control, whilst SMG7L has lost NMD activity and NMD control.
KeywordsNMD SMG7 Grapevine VIGS Gene duplication Plant
We are grateful to C. Lacomme (University of Edinburgh), D. Baulcombe (University of Cambridge) and SP. Dinesh-Kumar (Yale University) for TRV vectors. We thank Margaret Boulton (John Innes Centre, Norwich, UK) for language corrections. This research was supported by grants from the OTKA (K81481, CK80029). We thank E. Barta (University of Debrecen) for his help with bioinformatical studies. A. Hangyáné Benkovics is a graduate student of Corvinus University of Budapest (“Program of Viticulture” at the Doctoral School of Horticulture). T. Nyikó is a graduate student of ELTE “Classical and Molecular Genetics” PhD program.
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