Summary information and primary citation
- PDB-id
-
6d06;
DSSR-derived features in text and
JSON formats
- Class
- hydrolase-RNA
- Method
- X-ray (2.55 Å)
- Summary
- Human adar2d e488y mutant complexed with dsrna
containing an abasic site opposite the edited base
- Reference
-
Monteleone LR, Matthews MM, Palumbo CM, Thomas JM, Zheng
Y, Chiang Y, Fisher AJ, Beal PA (2019): "A
Bump-Hole Approach for Directed RNA Editing."
Cell Chem Biol, 26, 269. doi:
10.1016/j.chembiol.2018.10.025.
- Abstract
- Molecules capable of directing changes to nucleic acid
sequences are powerful tools for molecular biology and
promising candidates for the therapeutic correction of
disease-causing mutations. However, unwanted reactions at
off-target sites complicate their use. Here we report
selective combinations of mutant editing enzyme and
directing oligonucleotide. Mutations in human ADAR2
(adenosine deaminase acting on RNA 2) that introduce
aromatic amino acids at position 488 reduce background RNA
editing. This residue is juxtaposed to the nucleobase that
pairs with the editing site adenine, suggesting a steric
clash for the bulky mutants. Replacing this nucleobase with
a hydrogen atom removes the clash and restores editing
activity. A crystal structure of the E488Y mutant bound to
abasic site-containing RNA shows the accommodation of the
tyrosine side chain. Finally, we demonstrate directed RNA
editing in vitro and in human cells using mutant ADAR2
proteins and modified guide RNAs with reduced off-target
activity.
