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http://dx.doi.org/10.14348/molcells.2022.0130

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

Jun Seop, Yun (Department of Oral Pathology, Yonsei University College of Dentistry)
Hyeeun, Song (Department of Oral Pathology, Yonsei University College of Dentistry)
Nam Hee, Kim (Department of Oral Pathology, Yonsei University College of Dentistry)
So Young, Cha (Department of Oral Pathology, Yonsei University College of Dentistry)
Kyu Ho, Hwang (Department of Oral Pathology, Yonsei University College of Dentistry)
Jae Eun, Lee (Department of Oral Pathology, Yonsei University College of Dentistry)
Cheol-Hee, Jeong (Department of Oral Pathology, Yonsei University College of Dentistry)
Sang Hyun, Song (Department of Oral Pathology, Yonsei University College of Dentistry)
Seonghun, Kim (Department of Oral Pathology, Yonsei University College of Dentistry)
Eunae Sandra, Cho (Department of Oral Pathology, Yonsei University College of Dentistry)
Hyun Sil, Kim (Department of Oral Pathology, Yonsei University College of Dentistry)
Jong In, Yook (Department of Oral Pathology, Yonsei University College of Dentistry)
Publication Information
Molecules and Cells / v.45, no.12, 2022 , pp. 911-922 More about this Journal
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.
Keywords
Axin; Delta and Omicron variants; glycogen synthase kinase-3; nucleocapsid; phosphorylation; severe acute respiratory syndrome coronavirus 2;
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