Structural Insights into RNA Recognition by the Alternate-Splicing Regulator CUG-Binding Protein 1

Teplova, M.; Song, Jikui; Gaw, Hai Yan; Teplov, A. A.; Patel, Dinshaw J.

doi:10.1016/j.str.2010.06.018

Cited by 72 publications

(84 citation statements)

References 41 publications

(72 reference statements)

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“…2D-G). These motifs resemble the consensus motifs for hnRNPL and CELF/ Cugbp1, respectively (Teplova et al 2010;Ray et al 2013). Motif analysis of clusters lacking the UGCAUG-related motif failed to produce significant or high-information content motifs.…”

Section: Rbfox2 Iclip Is Strongly Enriched Around (U)gcaug Motifsmentioning

confidence: 67%

Rbfox2 controls autoregulation in RNA-binding protein networks

Boutz²,

et al. 2014

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The tight regulation of splicing networks is critical for organismal development. To maintain robust splicing patterns, many splicing factors autoregulate their expression through alternative splicing-coupled nonsensemediated decay (AS-NMD). However, as negative autoregulation results in a self-limiting window of splicing factor expression, it is unknown how variations in steady-state protein levels can arise in different physiological contexts. Here, we demonstrate that Rbfox2 cross-regulates AS-NMD events within RNA-binding proteins to alter their expression. Using individual nucleotide-resolution cross-linking immunoprecipitation coupled to highthroughput sequencing (iCLIP) and mRNA sequencing, we identified >200 AS-NMD splicing events that are bound by Rbfox2 in mouse embryonic stem cells. These ''silent'' events are characterized by minimal apparent splicing changes but appreciable changes in gene expression upon Rbfox2 knockdown due to degradation of the NMDinducing isoform. Nearly 70 of these AS-NMD events fall within genes encoding RNA-binding proteins, many of which are autoregulated. As with the coding splicing events that we found to be regulated by Rbfox2, silent splicing events are evolutionarily conserved and frequently contain the Rbfox2 consensus UGCAUG. Our findings uncover an unexpectedly broad and multilayer regulatory network controlled by Rbfox2 and offer an explanation for how autoregulatory splicing networks are tuned.

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Section: Rbfox2 Iclip Is Strongly Enriched Around (U)gcaug Motifsmentioning

confidence: 67%

Rbfox2 controls autoregulation in RNA-binding protein networks

Boutz²,

et al. 2014

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“…Gel shift assay CUGBP1 protein was purified as previously reported [15]. The RNA oligonucleotide (5′-AUUUGUU-3′) was synthesised by GenScript (Nanjing, China).…”

Section: Methodsmentioning

confidence: 99%

“…It preferentially binds to GU-rich elements located in the 5′ and 3′ untranslated regions (UTRs), as well as in mRNA coding regions [15,16]. The association of CUGBP1 with its mRNA targets influences their alternative splicing, translation and turnover [17].…”

Section: Introductionmentioning

confidence: 99%

RNA-binding protein CUGBP1 regulates insulin secretion via activation of phosphodiesterase 3B in mice

Zhai

Yang

et al. 2016

Diabetologia

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Aims/hypothesis CUG-binding protein 1 (CUGBP1) is a multifunctional RNA-binding protein that regulates RNA processing at several stages including translation, deadenylation and alternative splicing, as well as RNA stability. Recent studies indicate that CUGBP1 may play a role in metabolic disorders. Our objective was to examine its role in endocrine pancreas function through gain-and loss-of-function experiments and to further decipher the underlying molecular mechanisms. Methods A mouse model in which type 2 diabetes was induced by a high-fat diet (HFD; 60% energy from fat) and mice on a standard chow diet (10% energy from fat) were compared. Pancreas-specific CUGBP1 overexpression and knockdown mice were generated. Different lengths of the phosphodiesterase subtype 3B (PDE3B) 3′ untranslated region (UTR) were cloned for luciferase reporter analysis. Purified CUGBP1 protein was used for gel shift experiments.Results CUGBP1 is present in rodent islets and in beta cell lines; it is overexpressed in the islets of diabetic mice. Compared with control mice, the plasma insulin level after a glucose load was significantly lower and glucose clearance was greatly delayed in mice with pancreas-specific CUGBP1 overexpression; the opposite results were obtained upon pancreas-specific CUGBP1 knockdown. Glucose-and glucagon-like peptide1 (GLP-1)-stimulated insulin secretion was significantly attenuated in mouse islets upon CUGBP1 overexpression. This was associated with a strong decrease in intracellular cAMP levels, pointing to a potential role for cAMP PDEs. CUGBP1 overexpression had no effect on the mRNA levels of PDE1A, 1C, 2A, 3A, 4A, 4B, 4D, 7A and 8B subtypes, but resulted in increased PDE3B expression. CUGBP1 was found to directly bind to a specific ATTTGTT sequence residing in the 3′ UTR of PDE3B and stabilised PDE3B mRNA. In the presence of the PDE3 inhibitor cilostamide, glucose-and GLP-1-stimulated insulin secretion was no longer reduced by CUGBP1 overexpression. Similar to CUGBP1, PDE3B was overexpressed in the islets of diabetic mice. Conclusions/interpretation We conclude that CUGBP1 is a critical regulator of insulin secretion via activating PDE3B. Repressing this protein might provide a potential strategy for treating type 2 diabetes.

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“…The remaining bases have no counterpart in the SUP-12:RNA complex. The first two RRM domains of CUG-BP1 have also been characterized (PDB 3NNC) 17 . In the crystal structure with UUGUGUG, only the second RRM is found in complex with the RNA and once again the similarities are restricted to the central UG dinucleotide.…”

Section: Discussionmentioning

confidence: 99%

Backbone-independent nucleic acid binding by splicing factor SUP-12 reveals key aspects of molecular recognition

et al. 2014

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Cellular differentiation is frequently accompanied by alternative splicing, enabled by the expression of tissue-specific factors which bind to pre-mRNAs and regulate exon choice. During Caenorhabditis elegans development, muscle-specific expression of the splicing factor SUP-12, together with a member of the Fox-1 family of splicing proteins, generates a functionally distinct isoform of the fibroblast growth factor receptor EGL-15. Using a combination of NMR spectroscopy and isothermal titration calorimetry, we determined the mode of nucleic acid binding by the RNA recognition motif domain of SUP-12. The calculated structures provide the first atomic details of RNA and DNA binding by the family of proteins that include SUP-12, RBM24, RBM38/RNPC1, SEB-4 and XSeb4R. This information was further used to design strategic mutations to probe the interaction with ASD-1 and to quantitatively perturb splicing in vivo.

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Structural Insights into RNA Recognition by the Alternate-Splicing Regulator CUG-Binding Protein 1

Cited by 72 publications

References 41 publications

Rbfox2 controls autoregulation in RNA-binding protein networks

Rbfox2 controls autoregulation in RNA-binding protein networks

RNA-binding protein CUGBP1 regulates insulin secretion via activation of phosphodiesterase 3B in mice

Backbone-independent nucleic acid binding by splicing factor SUP-12 reveals key aspects of molecular recognition

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