Human 14-3-3 Proteins Site-selectively Bind the Mutational Hotspot Region of SARS-CoV-2 Nucleoprotein Modulating its Phosphoregulation.

Autor: Tugaeva KV; A.N. Bach Institute of Biochemistry, Federal Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia., Sysoev AA; A.N. Bach Institute of Biochemistry, Federal Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia., Kapitonova AA; A.N. Bach Institute of Biochemistry, Federal Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia., Smith JLR; York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5DD, United Kingdom., Zhu P; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA., Cooley RB; Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA., Antson AA; York Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5DD, United Kingdom., Sluchanko NN; A.N. Bach Institute of Biochemistry, Federal Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia. Electronic address: nikolai.sluchanko@mail.ru.
Jazyk: angličtina
Zdroj: Journal of molecular biology [J Mol Biol] 2023 Jan 30; Vol. 435 (2), pp. 167891. Date of Electronic Publication: 2022 Nov 24.
DOI: 10.1016/j.jmb.2022.167891
Abstrakt: Phosphorylation of SARS-CoV-2 nucleoprotein recruits human cytosolic 14-3-3 proteins playing a well-recognized role in replication of many viruses. Here we use genetic code expansion to demonstrate that 14-3-3 binding is triggered by phosphorylation of SARS-CoV-2 nucleoprotein at either of two pseudo-repeats centered at Ser197 and Thr205. According to fluorescence anisotropy measurements, the pT205-motif,presentin SARS-CoV-2 but not in SARS-CoV, is preferred over the pS197-motif by all seven human 14-3-3 isoforms, which collectively display an unforeseen pT205/pS197 peptide binding selectivity hierarchy. Crystal structures demonstrate that pS197 and pT205 are mutually exclusive 14-3-3-binding sites, whereas SAXS and biochemical data obtained on the full protein-protein complex indicate that 14-3-3 binding occludes the Ser/Arg-rich region of the nucleoprotein, inhibiting its dephosphorylation. This Ser/Arg-rich region is highly prone to mutations, as exemplified by the Omicron and Delta variants, with our data suggesting that the strength of 14-3-3/nucleoprotein interaction can be linked with the replicative fitness of the virus.
Competing Interests: Declaration of competing interests The authors declare no conflicts of interest.
(Copyright © 2022 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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