Poster abstracts
Poster number 65 submitted by Boyoon Yang
Exploring the importance of sequence-specific dsRNA recognition of ADARs in vivo
Boyoon Yang (Biochemistry Graduate Program, Indiana University, Bloomington, IN, USA), Ryan J. Andrews (Department of Biochemistry, University of Utah, Salt Lake City, UT, USA), Douglas B. Rusch (Center for Genomics and Bioinformatics, Indiana University, Bloomington, IN, USA), Brenda L. Bass (Department of Biochemistry, University of Utah, Salt Lake City, UT, USA), Heather A. Hundley (Department of Biology, Indiana University, Bloomington, IN, USA)
Abstract:
While recognition of double-stranded RNA (dsRNA) is known to be largely structure-specific rather than sequence-specific in vitro, it remains unclear how dsRNA binding proteins (dsRBPs) bind specific targets in vivo. Herein, we used eCLIP to identify in vivo binding sites of ADR-1 and ADR-2, members of the adenosine deaminase acting on RNA (ADAR) family of dsRBPs in Caenorhabditis elegans. We demonstrate that binding sites of the editing-deficient ADR-1 exhibit higher fold enrichment than the binding sites of the editing enzyme ADR-2, although ADR-1 is ~4-5 times less expressed at the protein level compared to ADR-2. Furthermore, ADR-1 binding sites exhibit narrower width than ADR-2, suggesting that ADR-1 binding events are more specific to certain regions. To explore how ADR-1 confers specific binding, we sought to test if ADR-1 binding sites are enriched for specific sequences. We revealed that a subset of ADR-1 binding sites is enriched for a 20-nt RNA sequence motif and tested if ADR-1 binding is affected in vivo. Our study provides new insights into how dsRBPs recognize specific targets and highlights the importance of an RNA sequence motif in dsRNA binding.
Keywords: RNA editing, ADAR, Substrate specificity