Structural and hydrodynamic characterization of dimeric human oligoadenylate synthetase 2
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Date
2020
Authors
Koul, Amit
Gemmill, Darren
Lubna, Nikhat
Meier, Markus
Krahn, Natalie
Booy, Evan P.
Stetefeld, Jörg
Patel, Trushar R.
McKenna, Sean A.
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Volume Title
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Abstract
Oligoadenylate synthetases (OASs) are a family of interferon-inducible enzymes that require double-stranded RNA (dsRNA) as a cofactor. Upon binding dsRNA, OAS undergoes a conformational change and is activated to polymerize ATP into 2′-5′-oligoadenylate chains. The OAS family consists of several isozymes, with unique domain organizations to potentially interact with dsRNA of variable length, providing diversity in viral RNA recognition. In addition, oligomerization of OAS isozymes, potentially OAS1 and OAS2, is hypothesized to be important for 2′-5′-oligoadenylate chain building. In this study, we present the solution conformation of dimeric human OAS2 using an integrated approach involving small-angle x-ray scattering, analytical ultracentrifugation, and dynamic light scattering techniques. We also demonstrate OAS2 dimerization using immunoprecipitation approaches in human cells. Whereas mutation of a key active-site aspartic acid residue prevents OAS2 activity, a C-terminal mutation previously hypothesized to disrupt OAS self-association had only a minor effect on OAS2 activity. Finally, we also present the solution structure of OAS1 monomer and dimer, comparing their hydrodynamic properties with OAS2. In summary, our work presents the first, to our knowledge, dimeric structural models of OAS2 that enhance our understanding of the oligomerization and catalytic function of OAS enzymes.
Description
Open access article. Creative Commons Attribution 4.0 International license (CC BY 4.0) applies
Keywords
Oligoadenylate synthetases , Dimeric structure , Hydrodynamic studies , OAS enzymes
Citation
Koul, A., Gemmill, D., Lubna, N., Meier, M., Krahn, N., Booy, E. P., Stetefeld, J., Patel, T. R., & McKenna, S. A. (2020). Structural and hydrodynamic characterization of dimeric human oligoadenylate synthetase 2. Biophysical Journal, 118(11), 2726-2740. https://doi.org/10.1016/j.bpj.2020.04.025