Roquin Promotes Constitutive mRNA Decay via a Conserved Class of Stem-Loop Recognition Motifs

Leppek, Kathrin; Schott, Johanna; Reitter, Sonja; Poetz, Fabian; Hammond, Ming C.; Stoecklin, Georg

doi:10.1016/j.cell.2013.04.016

Cited by 280 publications

(431 citation statements)

References 43 publications

Supporting

Mentioning

401

Contrasting

Unclassified

Order By: Relevance

“…Wild-type RC3H1 2-452 binds with a K d value of 139±42 nM to ICOS stem-loop mRNA (Fig. 2a), which is comparable with the binding constant reported for another CDE-containing stem-loop mRNA, namely TNF-a (92 ± 14 nM) 1 . On the other hand, RC3H1 2-452 variants with single basic patch mutations (RK219AA or KR259AA, respectively) showed significantly reduced ICOS binding while simultaneous mutation of the two basic patches nearly abolished binding ( Fig.…”

Section: Resultssupporting

confidence: 83%

“…It is conceivable that in addition to the reported CDE-containing stem-loop mRNAs also other structure-sequence motifs are recognized within targeted mRNAs, as also indicated by the fact that the majority of the mRNAs enriched in RC3H1 immunoprecipitation experiments were lacking the CDE motif 1 . This opens up exciting prospects for future research, addressing further biological roles of Roquin family members.…”

Section: Discussionmentioning

confidence: 99%

“…Roquin proteins, for example, were recently reported to specifically bind to a newly discovered conserved class of RNA stem-loop motifs, known as constitutive decay elements (CDE), which are present in the 3 0 UTR of various mRNAs, thereby initiating their degradation. On a transcriptome-wide scale, 450 vertebrate CDEcontaining mRNAs were identified as Roquin targets, many of which encode proteins important for development as well as inflammation and immunity 1 . Roquin binding to the CDE in the 3 0 UTR of tumour necrosis factor-a (TNF-a) mRNA, for example, limits the synthesis of the proinflammatory cytokine TNF-a (ref.…”

mentioning

confidence: 99%

“…Roquin binding to the CDE in the 3 0 UTR of tumour necrosis factor-a (TNF-a) mRNA, for example, limits the synthesis of the proinflammatory cytokine TNF-a (ref. 1). The expression of ICOS (inducible T-cell costimulator), a gene encoding an essential receptor for follicular T-cells, is also restricted by Roquin, which promotes the degradation of ICOS mRNA, thereby playing an important role in autoimmunity 2,3 .…”

mentioning

confidence: 99%

“…Human ROQUIN1, also known as RC3H1 (ring finger and CCCH-type zinc finger domains 1), is a 1,133 amino-acid protein, containing a RING (really interesting new gene)-type zinc finger, a roquin domain (ROQ), a CCCH-type zinc finger (zf), and a poorly defined carboxy (C)-terminal, proline-rich domain. Murine Roquin binding to CDE-like RNA stem-loop motifs was shown to be mediated by the ROQ domain, while the C-terminal effector domain recruits the Ccr4-Caf1-Not complex, resulting in Caf1a-dependent deadenylation and consecutive degradation of CDEcontaining mRNAs 1 . Although Roquin proteins were shown to act broadly as mediators of mRNA deadenylation by recognizing a conserved class of stem-loop RNA degradation motifs via the ROQ domain 1 , the structural basis for stem-loop mRNA recognition is still unknown.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Roquin binding to target mRNAs involves a winged helix-turn-helix motif

et al. 2014

View full text Add to dashboard Cite

Roquin proteins mediate mRNA deadenylation by recognizing a conserved class of stem-loop RNA degradation motifs via their Roquin domain. Here we present the crystal structure of a Roquin domain, revealing a mostly helical protein fold bearing a winged helix-turn-helix motif. By combining structural, biochemical and mutation analyses, we gain insight into the mode of RNA binding. We show that the winged helix-turn-helix motif is involved in the binding of constitutive decay elements-containing stem-loop mRNAs. Moreover, we provide biochemical evidence that Roquin proteins are additionally able to bind to duplex RNA and have the potential to be functional in different oligomeric states.

show abstract

Section: Resultssupporting

confidence: 83%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Roquin binding to target mRNAs involves a winged helix-turn-helix motif

et al. 2014

View full text Add to dashboard Cite

show abstract

Exaptive origins of regulated mRNA decay in eukaryotes

Hamid

Makeyev

2016

BioEssays

View full text Add to dashboard Cite

Eukaryotic gene expression is extensively controlled at the level of mRNA stability and the mechanisms underlying this regulation are markedly different from their archaeal and bacterial counterparts. We propose that two such mechanisms, nonsense‐mediated decay (NMD) and motif‐specific transcript destabilization by CCCH‐type zinc finger RNA‐binding proteins, originated as a part of cellular defense against RNA pathogens. These branches of the mRNA turnover pathway might have been used by primeval eukaryotes alongside RNA interference to distinguish their own messages from those of RNA viruses and retrotransposable elements. We further hypothesize that the subsequent advent of “professional” innate and adaptive immunity systems allowed NMD and the motif‐triggered mechanisms to be efficiently repurposed for regulation of endogenous cellular transcripts. This scenario explains the rapid emergence of archetypical mRNA destabilization pathways in eukaryotes and argues that other aspects of post‐transcriptional gene regulation in this lineage might have been derived through a similar exaptation route.

show abstract

Discovery of Small‐Molecule Modulators of Protein–RNA Interactions by Fluorescence Intensity‐Based Binding Assay

2019

View full text Add to dashboard Cite

Protein–RNA interactions mediate various cellular processes, the dysregulation of which has been associated with a list of diseases. Thus, novel experimental tools for monitoring protein–RNA interactions are highly desirable to identify new chemical modulators of these therapeutic targets. In this study, we constructed simple fluorescence intensity‐based protein–RNA binding assays by testing multiple environment‐sensitive organic fluorophores. We selected the oncogenic interaction between Lin28 and the let‐7 microRNA and the important immunomodulatory Roquin–Tnf CDE interaction as representative targets. We adapted this assay to high‐throughput screening for the identification of pyrazolyl thiazolidinedione‐type molecules as potent small‐molecule inhibitors of protein–microRNA interactions. We clearly showed the structure–activity relationships of this new class of Lin28–let‐7 interaction inhibitors, and confirmed that cellular mature let‐7 microRNAs and their target genes could be modulated upon treatment with the pyrazolyl thiazolidinedione‐type inhibitor. We expect that our simple and adaptable screening approach can be applied for the development of various assay systems aimed at the identification of bioactive small molecules targeting protein–RNA interactions.

show abstract

Roquin Promotes Constitutive mRNA Decay via a Conserved Class of Stem-Loop Recognition Motifs

Cited by 280 publications

References 43 publications

Roquin binding to target mRNAs involves a winged helix-turn-helix motif

Roquin binding to target mRNAs involves a winged helix-turn-helix motif

Exaptive origins of regulated mRNA decay in eukaryotes

Discovery of Small‐Molecule Modulators of Protein–RNA Interactions by Fluorescence Intensity‐Based Binding Assay

Contact Info

Product

Resources

About