1. G3BP1 binds to guanine quadruplexes in mRNAs to modulate their stabilities.
- Author
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He, Xiaomei, Yuan, Jun, and Wang, Yinsheng
- Subjects
3 Untranslated Regions ,Aminoquinolines ,Base Sequence ,Cloning ,Molecular ,Computational Biology ,DNA Helicases ,Datasets as Topic ,Escherichia coli ,G-Quadruplexes ,Gene Expression ,Genes ,Reporter ,Genetic Vectors ,HEK293 Cells ,HeLa Cells ,Humans ,Ligands ,Luciferases ,Picolinic Acids ,Poly-ADP-Ribose Binding Proteins ,Protein Binding ,RNA Helicases ,RNA Recognition Motif Proteins ,RNA Stability ,Recombinant Proteins - Abstract
RNA guanine quadruplexes (rG4) assume important roles in post-transcriptional regulations of gene expression, which are often modulated by rG4-binding proteins. Hence, understanding the biological functions of rG4s requires the identification and functional characterizations of rG4-recognition proteins. By employing a bioinformatic approach based on the analysis of overlap between peaks obtained from rG4-seq analysis and those detected in >230 eCLIP-seq datasets for RNA-binding proteins generated from the ENCODE project, we identified a large number of candidate rG4-binding proteins. We showed that one of these proteins, G3BP1, is able to bind directly to rG4 structures with high affinity and selectivity, where the binding entails its C-terminal RGG domain and is further enhanced by its RRM domain. Additionally, our seCLIP-Seq data revealed that pyridostatin, a small-molecule rG4 ligand, could displace G3BP1 from mRNA in cells, with the most pronounced effects being observed for the 3-untranslated regions (3-UTR) of mRNAs. Moreover, luciferase reporter assay results showed that G3BP1 positively regulates mRNA stability through its binding with rG4 structures. Together, we identified a number of candidate rG4-binding proteins and validated that G3BP1 can bind directly with rG4 structures and regulate the stabilities of mRNAs.
- Published
- 2021