Staufen2, like Staufen1, functions in SMD by binding to itself and its paralog and promoting UPF1 helicase but not ATPase activity
ABSTRACT Staufen (STAU)1-mediated mRNA decay (SMD) is a posttranscriptional regulatory mechanism in mammals that degrades mRNAs harboring a STAU1-binding site (SBS) in their 3'-untranslated regions (3' UTRs). We show that SMD involves not only STAU1 but also its paralog STAU2. STAU2, like STAU1, is a double-stranded RNA-binding protein that interacts directly with the ATP-dependent RNA helicase up-frameshift 1 (UPF1) to reduce the half-life of SMD targets that form an SBS by either intramolecular or intermolecular base-pairing. Compared with STAU1, STAU2 binds ∼10-fold more UPF1 and ∼two- to fivefold more of those SBS-containing mRNAs that were tested, and it comparably promotes UPF1 helicase activity, which is critical for SMD. STAU1- or STAU2-mediated augmentation of UPF1 helicase activity is not accompanied by enhanced ATP hydrolysis but does depend on ATP binding and a basal level of UPF1 ATPase activity. Studies of STAU2 demonstrate it changes the conformation of RNA-bound UPF1. These findings, and evidence for STAU1-STAU1, STAU2-STAU2, and STAU1-STAU2 formation in vitro and in cells, are consistent with results from tethering assays: the decrease in mRNA abundance brought about by tethering siRNA-resistant STAU2 or STAU1 to an mRNA 3' UTR is inhibited by downregulating the abundance of cellular STAU2, STAU1, or UPF1. It follows that the efficiency of SMD in different cell types reflects the cumulative abundance of STAU1 and STAU2. We propose that STAU paralogs contribute to SMD by "greasing the wheels" of RNA-bound UPF1 so as to enhance its unwinding capacity per molecule of ATP hydrolyzed.
- SourceAvailable from: Osamu Takeuchi
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- "Whereas all three UPF members are required for NMD, UPF2 and UPF3x are also dispensable for SMD and replication-dependent histone mRNA decay (Kaygun and Marzluff, 2005; Kim et al., 2005). Nevertheless, Staufen 1 and 2 are sensors for recognizing dsRNA sequences (Park et al., 2013), but do not have RNase activity. "
ABSTRACT: Regnase-1 and Roquin are RNA binding proteins essential for degradation of inflammation-related mRNAs and maintenance of immune homeostasis. However, their mechanistic relationship has yet to be clarified. Here, we show that, although Regnase-1 and Roquin regulate an overlapping set of mRNAs via a common stem-loop structure, they function in distinct subcellular locations: ribosome/endoplasmic reticulum and processing-body/stress granules, respectively. Moreover, Regnase-1 specifically cleaves and degrades translationally active mRNAs and requires the helicase activity of UPF1, similar to the decay mechanisms of nonsense mRNAs. In contrast, Roquin controls translationally inactive mRNAs, independent of UPF1. Defects in both Regnase-1 and Roquin lead to large increases in their target mRNAs, although Regnase-1 tends to control the early phase of inflammation when mRNAs are more actively translated. Our findings reveal that differential regulation of mRNAs by Regnase-1 and Roquin depends on their translation status and enables elaborate control of inflammation. Copyright © 2015 Elsevier Inc. All rights reserved.Cell 05/2015; 161(5):1058-1073. DOI:10.1016/j.cell.2015.04.029 · 33.12 Impact Factor
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- "First, we have provided evidence for a role of Stau2 in the stabilization of mRNAs as Stau2 targets were predominantly downregulated in Stau2-deficient neurons. We note that the levels of a small fraction of Stau2 targets increase upon Stau2 downregulation, consistent with a recent study in human cell lines that implicates Stau2 in transcript destabilization (Park et al., 2013). Although no such role of transcript stabilization has been reported for Stau2 before, there is a recent publication for Stau1, together with the long noncoding RNA TINCR, showing a role in stabilizing differentiation mRNAs in human keratinocytes (Kretz et al., 2013). "
ABSTRACT: RNA-binding proteins play crucial roles in directing RNA translation to neuronal synapses. Staufen2 (Stau2) has been implicated in both dendritic RNA localization and synaptic plasticity in mammalian neurons. Here, we report the identification of functionally relevant Stau2 target mRNAs in neurons. The majority of Stau2-copurifying mRNAs expressed in the hippocampus are present in neuronal processes, further implicating Stau2 in dendritic mRNA regulation. Stau2 targets are enriched for secondary structures similar to those identified in the 3' UTRs of Drosophila Staufen targets. Next, we show that Stau2 regulates steady-state levels of many neuronal RNAs and that its targets are predominantly downregulated in Stau2-deficient neurons. Detailed analysis confirms that Stau2 stabilizes the expression of one synaptic signaling component, the regulator of G protein signaling 4 (Rgs4) mRNA, via its 3' UTR. This study defines the global impact of Stau2 on mRNAs in neurons, revealing a role in stabilization of the levels of synaptic targets.Cell Reports 12/2013; 5(6). DOI:10.1016/j.celrep.2013.11.039 · 8.36 Impact Factor
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- "Two days after siRNA transfection, cells were re-transfected with a pmCMV-Gl (β-globin) test plasmid , either nonsense-free (Norm) or PTC-harboring (Ter), a pmCMV-GPx1 (glutathione peroxidase 1) test plasmid , and a phCMV-MUP (mouse major urinary protein) reference plasmid . When specified, pCI-Neo–FLAG–UPF1  was also transfected. Total-cell lysates were obtained by hypotonic solution containing 10 mM Tris–Cl pH 7.5, 10 mM NaCl, 10 mM EDTA, and 0.5% Triton X-100 followed by a 10-min incubation on ice as done in the previous report . "
ABSTRACT: Nonsense-mediated mRNA decay (NMD) is the best-characterized mRNA surveillance mechanism that degrades a premature-termination codon (PTC)-containing mRNA. During mammalian NMD, SMG1 and UPF1, key proteins in NMD, join at a PTC and form an SMG1-UPF1-eRF1-eRF3 (SURF) complex by binding UPF1 to eRF3 after PTC-recognition by the translating ribosome. Subsequently, UPF1 is phosphorylated after UPF1-SMG1 moves onto the downstream exon junction complex (EJC). However, the cellular events that induce UPF1 and SMG1 complex formation and increase NMD efficiency before PTC recognition remain unclear. Here, we show that telomere-maintenance 2 (TEL2) phosphorylation by casein-kinase 2 (CK2) increases SMG1 stability, which increases UPF1 phosphorylation and, ultimately, augments NMD. Inhibition of CK2 activity or downregulation of TEL2 impairs NMD. Intriguingly, loss of TEL2 phosphorylation reduces UPF1-bound PTC-containing mRNA and the formation of the SMG1-UPF1 complex. Thus, our results identify a new function of CK2-mediated TEL2 phosphorylation in a mammalian NMD.Biochimica et Biophysica Acta 07/2013; 1829(10). DOI:10.1016/j.bbagrm.2013.06.002 · 4.66 Impact Factor