CK2 phosphorylation of human papillomavirus 16 E2 on serine 23 promotes interaction with TopBP1 and Is critical for E2 interaction with mitotic chromatin and the viral life cycle

Prabhakar, Apurva T, James, Claire D, Das, Dipon, Otoa, Raymonde, Day, Matthew, Burgner, John, Fontan, Christian T, Wang, Xu, Glass, Sarah H, Wieland, Andreas, Donaldson, Mary M, Bristol, Molly L, Li, Renfeng, Oliver, Antony W, Pearl, Laurence H, Smith, Brian O and Morgana, Iain M (2021) CK2 phosphorylation of human papillomavirus 16 E2 on serine 23 promotes interaction with TopBP1 and Is critical for E2 interaction with mitotic chromatin and the viral life cycle. mBio, 12 (5). pp. 1-23. ISSN 2150-7511

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Abstract

During the human papillomavirus 16 (HPV16) life cycle, the E2 protein interacts with host factors to regulate viral transcription, replication, and genome segregation/retention. Our understanding of host partner proteins and their roles in E2 functions remains incomplete. Here we demonstrate that CK2 phosphorylation of E2 on serine 23 promotes interaction with TopBP1 in vitro and in vivo and that E2 is phosphorylated on this residue during the HPV16 life cycle. We investigated the consequences of mutating serine 23 on E2 functions. E2-S23A (E2 with serine 23 mutated to alanine) activates and represses transcription identically to E2-WT (wild-type E2), and E2-S23A is as efficient as E2-WT in transient replication assays. However, E2-S23A has compromised interaction with mitotic chromatin compared with E2-WT. In E2-WT cells, both E2 and TopBP1 levels increase during mitosis compared with vector control cells. In E2-S23A cells, neither E2 nor TopBP1 levels increase during mitosis. Introduction of the S23A mutation into the HPV16 genome resulted in delayed immortalization of human foreskin keratinocytes (HFK) and higher episomal viral genome copy number in resulting established HFK. Remarkably, S23A cells had a disrupted viral life cycle in organotypic raft cultures, with a loss of E2 expression and a failure of viral replication. Overall, our results demonstrate that CK2 phosphorylation of E2 on serine 23 promotes interaction with TopBP1 and that this interaction is critical for the viral life cycle.

Item Type: Article
Keywords: BRD4, CK2, TopBP1, E2, cervical cancer, head and neck cancer, human papillomavirus
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 12 Oct 2021 07:08
Last Modified: 02 Mar 2022 12:58
URI: http://sro.sussex.ac.uk/id/eprint/102254

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Structural Biology of DNA Damage Response and Repair MechanismsG2176CANCER RESEARCH UKC302/A24386