Asymmetric dimerization of adenosine deaminase acting on RNA facilitates substrate recognition

Thuy-Boun, Alexander S.; Thomas, Justin M.; Grajo, Herra L.; Palumbo, Cody M.; Park, SeHee; Nguyen, Luan T.; Fisher, Andrew J.; Beal, Peter A.

doi:10.1093/nar/gkaa532

Asymmetric dimerization of adenosine deaminase acting on RNA facilitates substrate recognition

Journal Article · Mon Jun 29 00:00:00 EDT 2020 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkaa532· OSTI ID:1635307

^[1]; Thomas, Justin M. ^[1]; Grajo, Herra L. ^[1]; Palumbo, Cody M. ^[1]; Park, SeHee ^[1]; Nguyen, Luan T. ^[1]; ^[2]; ^[1]

Department of Chemistry, University of California, Davis, CA, USA
Department of Chemistry, University of California, Davis, CA, USA, Department of Molecular and Cellular Biology, University of California, Davis, CA, USA

Abstract

Adenosine deaminases acting on RNA (ADARs) are enzymes that convert adenosine to inosine in duplex RNA, a modification that exhibits a multitude of effects on RNA structure and function. Recent studies have identified ADAR1 as a potential cancer therapeutic target. ADARs are also important in the development of directed RNA editing therapeutics. A comprehensive understanding of the molecular mechanism of the ADAR reaction will advance efforts to develop ADAR inhibitors and new tools for directed RNA editing. Here we report the X-ray crystal structure of a fragment of human ADAR2 comprising its deaminase domain and double stranded RNA binding domain 2 (dsRBD2) bound to an RNA duplex as an asymmetric homodimer. We identified a highly conserved ADAR dimerization interface and validated the importance of these sequence elements on dimer formation via gel mobility shift assays and size exclusion chromatography. We also show that mutation in the dimerization interface inhibits editing in an RNA substrate-dependent manner for both ADAR1 and ADAR2.

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Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1635307

Alternate ID(s):: OSTI ID: 2471721

Journal Information:: Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 14 Vol. 48; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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