Abstract
Adenosine-to-inosine (A-to-I) editing of a subset of RNAs in a eukaryotic cell is required in order to avoid triggering the innate immune system. Editing is carried out by ADAR1, which exists as short (p110) and long (p150) isoforms. ADAR1p150 is mostly cytoplasmic, possesses a Z-RNA binding domain (Zα), and is only expressed during the innate immune response. A structurally homologous domain to Zα, the Zβ domain, is separated by a long linker from Zα on the N-terminus of ADAR1 but its function remains unknown. Zβ does not bind to RNA in isolation, yet the binding kinetics of the segment encompassing Zα, Zβ and the 95-residue linker between the two domains (Zα–Zβ) are markedly different compared to Zα alone. Here we present the solution NMR backbone assignment of Zα–Zβ from H. Sapiens ADAR1. The predicted secondary structure of Zα–Zβ based on chemical shifts is in agreement with previously determined structures of Zα and Zβ in isolation, and indicates that the linker is intrinsically disordered. Comparison of the chemical shifts between the individual Zα and Zβ domains to the full Zα–Zβ construct suggests that Zβ may interact with the linker, the function of which is currently unknown.
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Data availability
The chemical shift assignments for Zα (BMRB 50714), Zβ (BMRB 50713) and Zα–Zβ (BMRB 50715) have been deposited in the Biological Magnetic Resonance Data Bank.
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Acknowledgements
The authors thank David Jones (University of Colorado, Aurora) for help with NMR spectroscopy, and Jeffrey Kieft for support. This project was supported by NIH Grant R01GM130694-01A1, NSF Grant 1917254 for Infrastructure Innovation for Biological Research, and a start-up package by the University of Colorado to B.V., and University of Colorado Cancer Center Support Grant P30 CA046934 and NIH Biomedical Research Support Shared Grant S10 OD025020-01.
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Nichols, P.J., Henen, M.A., Vicens, Q. et al. Solution NMR backbone assignments of the N-terminal Zα-linker-Zβ segment from Homo sapiens ADAR1p150. Biomol NMR Assign 15, 273–279 (2021). https://doi.org/10.1007/s12104-021-10017-8
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DOI: https://doi.org/10.1007/s12104-021-10017-8