Structure-function studies of STAR family quaking proteins bound to their in vivo RNA target sites Journal Article
| Authors: | Teplova, M.; Hafner, M.; Teplov, D.; Essig, K.; Tuschl, T.; Patel, D. J. |
| Article Title: | Structure-function studies of STAR family quaking proteins bound to their in vivo RNA target sites |
| Abstract: | Mammalian Quaking (QKI) and its Caenorhabditis elegans homolog, GLD-1 (defective in germ line development), are evolutionarily conserved RNA-binding proteins, which post-transcriptionally regulate target genes essential for developmental processes and myelination. We present X-ray structures of the STAR (signal transduction and activation of RNA) domain, composed of Qua1, K homology (KH), and Qua2 motifs of QKI and GLD-1 bound to high-affinity in vivo RNA targets containing YUAAY RNA recognition elements (RREs). The KH and Qua2 motifs of the STAR domain synergize to specifically interact with bases and sugar-phosphate backbones of the bound RRE. Qua1-mediated homodimerization generates a scaffold that enables concurrent recognition of two RREs, thereby plausibly targeting tandem RREs present in many QKI-targeted transcripts. Structure-guided mutations reduced QKI RNA-binding affinity in vitro and in vivo, and expression of QKI mutants in human embryonic kidney cells (HEK293) significantly decreased the abundance of QKI target mRNAs. Overall, our studies define principles underlying RNA target selection by STAR homodimers and provide insights into the post-transcriptional regulatory function of mammalian QKI proteins. © 2013 by Cold Spring Harbor Laboratory Press. |
| Keywords: | protein expression; unclassified drug; human cell; mutation; binding affinity; protein domain; protein function; protein motif; mammalia; animals; gene targeting; embryo; protein protein interaction; genetic transcription; in vivo study; in vitro study; rna binding protein; rna; gene expression regulation; rna-binding proteins; transcription regulation; amino acid sequence; molecular sequence data; cell strain hek293; messenger rna; sequence alignment; escherichia coli; binding site; crystal structure; models, molecular; dimerization; protein structure, tertiary; caenorhabditis elegans; protein structure; x ray crystallography; protein structure, quaternary; myelination; x-ray crystallography; mrna; protein rna binding; caenorhabditis elegans protein; hek293 cells; caenorhabditis elegans proteins; scaffold protein; homodimer; gld-1; par-clip; qki; gld 1 protein; quaking protein; signal transduction and activation of rna domain protein; sugar phosphate |
| Journal Title: | Genes and Development |
| Volume: | 27 |
| Issue: | 8 |
| ISSN: | 0890-9369 |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Date Published: | 2013-04-15 |
| Start Page: | 928 |
| End Page: | 940 |
| Language: | English |
| DOI: | 10.1101/gad.216531.113 |
| PROVIDER: | scopus |
| PMCID: | PMC3650229 |
| PUBMED: | 23630077 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 3 June 2013" - "CODEN: GEDEE" - "Source: Scopus" |
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