8E0F

Human Adenosine Deaminase Acting on dsRNA (ADAR2-RD) bound to dsRNA containing a G-G pair adjacent to the target site

Summary for 8E0F

• Entry DOI: 10.2210/pdb8e0f/pdb
• Descriptor: Double-stranded RNA-specific editase 1, RNA (5-R(*GP*CP*UP*CP*GP*CP*GP*AP*UP*GP*CP*GP*(8AZ)P*GP*AP*GP*GP*GP*CP* UP*CP*UP*GP*AP*UP*AP*GP*CP*UP*AP*CP*G)-3), RNA(5-R(*CP*GP*UP*AP*GP*CP*UP*AP*UP*CP*AP*GP*AP*GP*CP*CP*CP*CP*CP*CP*GP*GP*CP*AP*UP*CP*GP*CP*GP*AP*GP*C)-3), ... (6 entities in total)
• Functional Keywords: protein-rna complex, rna editing, rna binding protein, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 130014.66

Authors

Wilcox, X.E.,Fisher, A.J.,Beal, P.A. (deposition date: 2022-08-09, release date: 2022-10-26, Last modification date: 2023-10-18)

Primary citation

Doherty, E.E.,Karki, A.,Wilcox, X.E.,Mendoza, H.G.,Manjunath, A.,Matos, V.J.,Fisher, A.J.,Beal, P.A.
ADAR activation by inducing a syn conformation at guanosine adjacent to an editing site.
Nucleic Acids Res., 50:10857-10868, 2022

PubMed Abstract: ADARs (adenosine deaminases acting on RNA) can be directed to sites in the transcriptome by complementary guide strands allowing for the correction of disease-causing mutations at the RNA level. However, ADARs show bias against editing adenosines with a guanosine 5' nearest neighbor (5'-GA sites), limiting the scope of this approach. Earlier studies suggested this effect arises from a clash in the RNA minor groove involving the 2-amino group of the guanosine adjacent to an editing site. Here we show that nucleosides capable of pairing with guanosine in a syn conformation enhance editing for 5'-GA sites. We describe the crystal structure of a fragment of human ADAR2 bound to RNA bearing a G:G pair adjacent to an editing site. The two guanosines form a Gsyn:Ganti pair solving the steric problem by flipping the 2-amino group of the guanosine adjacent to the editing site into the major groove. Also, duplexes with 2'-deoxyadenosine and 3-deaza-2'-deoxyadenosine displayed increased editing efficiency, suggesting the formation of a Gsyn:AH+anti pair. This was supported by X-ray crystallography of an ADAR complex with RNA bearing a G:3-deaza dA pair. This study shows how non-Watson-Crick pairing in duplex RNA can facilitate ADAR editing enabling the design of next generation guide strands for therapeutic RNA editing.

PubMed: 36243986
DOI: 10.1093/nar/gkac897

Experimental method

X-RAY DIFFRACTION (2.7 Å)