University of Colorado
Ozdilek, Bagdeser Akdogan.
Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains.
Degree: PhD, 2017, University of Colorado
Recent developments in the comprehensive identification of the RNA-binding protein (RBP) repertoire has accelerated discovery of new RBPs. According to these studies, 20% of both known and novel RNA-binding proteins are highly disordered. Moreover, analysis of the human mRNA interactome revealed most disease related mutations are found within intrinsically disordered RNA-binding domains (RBDs). For most of these proteins, their RNA-binding properties are poorly characterized. Thus, deciphering intrinsically disordered RBD-RNA interactions on a molecular scale is essential to understanding their impact upon human physiology and diseases.
RGG/RG (arginine/glycine) domains are the second most common RNA binding domain in the human genome, yet their RNA-binding properties have not been well understood. Proteins containing RGG/RG domains regulate all aspects of RNA metabolism including transcription, processing, nucleocytoplasmic shuttling and translation. Proteins such as Fused in Sarcoma (FUS), Fragile X mental retardation (FMRP) and hnRNP U, bind a majority of the cellular transcriptome such that their recognition of RNAs has been considered to be non-specific or “promiscuous”.
Here, I report a detailed analysis of the RNA-binding characteristics of the RGG/RG domains from FUS, FMRP and hnRNP U. While previous studies of FUS focused on RNA binding by the RRM and zinc finger (ZnF) domains, my analysis showed RNA binding activity is driven by the RGG/RG domains. Further, I observed a strong synergy between the RRM and adjacent RGG/RG domains to achieve RNA binding affinities of the full-length FUS. To better characterize RNA-binding properties of RGG/RG domains, we have analyzed RGG/RG domains of FUS, FMRP and hnRNP U in vitro
against a spectrum of different RNAs with well-defined structural and sequence features. These experiments revealed that RGG/RG domains have different degrees of preference for binding to RNAs but share consistent trends in their selectivity towards RNAs with complex secondary structure. Thus, the binding behavior of RGG domains is best described as “degenerate specificity” reflecting that RGG/RG domains interact with a broad spectrum of RNAs that contain frequently observed sequence/structural elements. This mode of specificity is likely further facilitated by the intrinsically disordered nature of RGG/RG domains that enable them to adopt multiple conformations to adaptively bind RNA.
Advisors/Committee Members: Robert T. Batey, Ravinder Singh, Deborah S. Wuttke, Shelley Copley, Thomas Perkins.
Subjects/Keywords: RNA-binding protein; RGG/RG domains; Biochemistry; Molecular Biology
to Zotero / EndNote / Reference
APA (6th Edition):
Ozdilek, B. A. (2017). Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains. (Doctoral Dissertation). University of Colorado. Retrieved from https://scholar.colorado.edu/mcdb_gradetds/77
Chicago Manual of Style (16th Edition):
Ozdilek, Bagdeser Akdogan. “Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains.” 2017. Doctoral Dissertation, University of Colorado. Accessed January 22, 2020.
MLA Handbook (7th Edition):
Ozdilek, Bagdeser Akdogan. “Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains.” 2017. Web. 22 Jan 2020.
Ozdilek BA. Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains. [Internet] [Doctoral dissertation]. University of Colorado; 2017. [cited 2020 Jan 22].
Available from: https://scholar.colorado.edu/mcdb_gradetds/77.
Council of Science Editors:
Ozdilek BA. Characterizing the RNA Binding Properties of the Intrinsically Disordered FUS Protein and RGG/RG Domains. [Doctoral Dissertation]. University of Colorado; 2017. Available from: https://scholar.colorado.edu/mcdb_gradetds/77