Glycogen Synthase Kinase-3 Interaction Domain Enhances Phosphorylation of SARS-CoV-2 Nucleocapsid Protein

Jun Seop Yun, Hyeeun Song, Nam Hee Kim, So Young Cha, Kyu Ho Hwang, Jae Eun Lee, Cheol Hee Jeong, Sang Hyun Song, Seonghun Kim, Eunae Sandra Cho, Hyun Sil Kim, Jong In Yook

Research output: Contribution to journalArticlepeer-review

Abstract

A structural protein of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), nucleocapsid (N) protein is phosphorylated by glycogen synthase kinase (GSK)-3 on the serine/arginine (SR) rich motif located in disordered regions. Although phosphorylation by GSK-3β constitutes a critical event for viral replication, the molecular mechanism underlying N phosphorylation is not well understood. In this study, we found the putative alpha-helix L/FxxxL/AxxRL motif known as the GSK-3 interacting domain (GID), found in many endogenous GSK-3β binding proteins, such as Axins, FRATs, WWOX, and GSKIP. Indeed, N interacts with GSK-3β similarly to Axin, and Leu to Glu substitution of the GID abolished the interaction, with loss of N phosphorylation. The N phosphorylation is also required for its structural loading in a virus-like particle (VLP). Compared to other coronaviruses, N of Sarbecovirus lineage including bat RaTG13 harbors a CDK1-primed phosphorylation site and Gly-rich linker for enhanced phosphorylation by GSK-3β. Furthermore, we found that the S202R mutant found in Delta and R203K/G204R mutant found in the Omicron variant allow increased abundance and hyper-phosphorylation of N. Our observations suggest that GID and mutations for increased phosphorylation in N may have contributed to the evolution of variants.

Original languageEnglish
Pages (from-to)911-922
Number of pages12
JournalMolecules and cells
Volume45
Issue number12
DOIs
Publication statusPublished - 2022 Dec 31

Bibliographical note

Publisher Copyright:
© The Korean Society for Molecular and Cellular Biology.

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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