Journal: Nat Commun / Year: 2016 Title: The RNA helicase DHX34 functions as a scaffold for SMG1-mediated UPF1 phosphorylation. Authors: Roberto Melero / Nele Hug / Andrés López-Perrote / Akio Yamashita / Javier F Cáceres / Oscar Llorca / Abstract: Nonsense-mediated decay (NMD) is a messenger RNA quality-control pathway triggered by SMG1-mediated phosphorylation of the NMD factor UPF1. In recent times, the RNA helicase DHX34 was found to ...Nonsense-mediated decay (NMD) is a messenger RNA quality-control pathway triggered by SMG1-mediated phosphorylation of the NMD factor UPF1. In recent times, the RNA helicase DHX34 was found to promote mRNP remodelling, leading to activation of NMD. Here we demonstrate the mechanism by which DHX34 functions in concert with SMG1. DHX34 comprises two distinct structural units, a core that binds UPF1 and a protruding carboxy-terminal domain (CTD) that binds the SMG1 kinase, as shown using truncated forms of DHX34 and electron microscopy of the SMG1-DHX34 complex. Truncation of the DHX34 CTD does not affect binding to UPF1; however, it compromises DHX34 binding to SMG1 to affect UPF1 phosphorylation and hence abrogate NMD. Altogether, these data suggest the existence of a complex comprising SMG1, UPF1 and DHX34, with DHX34 functioning as a scaffold for UPF1 and SMG1. This complex promotes UPF1 phosphorylation leading to functional NMD.
History
Deposition
Dec 11, 2015
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Header (metadata) release
Jan 13, 2016
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Map release
Jan 13, 2016
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Update
Feb 17, 2016
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Current status
Feb 17, 2016
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
pH: 7.5 Details: 10 mM HEPES-KOH, 150 mM NaCl, 20% (v/v) glycerol, 10 mM MgCl2
Staining
Type: NEGATIVE / Details: 1% uranyl formate
Grid
Details: 400 mesh grid with thin carbon support, glow discharged
Vitrification
Cryogen name: NONE / Instrument: OTHER
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Electron microscopy
Microscope
JEOL 1230
Alignment procedure
Legacy - Astigmatism: Objective lens astigmatism was corrected using a TVIPS F416 CMOS and the EM-MENU software (TVIPS)
Date
Feb 17, 2012
Image recording
Category: CCD / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Digitization - Sampling interval: 15.6 µm / Average electron dose: 15 e/Å2 Details: Using a TVIPS F416 CMOS and the EM-TOOLS software (TVIPS) Bits/pixel: 16
Electron beam
Acceleration voltage: 100 kV / Electron source: TUNGSTEN HAIRPIN
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