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, Diptikanta; Otoa, Raymonde; Day, Matthew; Burgner, John W.; Fontan, Christian T.; Wang, Xu; Glass, Sarah H; Wieland, Andreas; Donaldson, Mary; Bristol, Molly L.; Li, Renfeng; Oliver, Anthony W.; Pearl, Laurence H.; Smith, Brian; Morgan, Iain M.

doi:10.1128/mbio.01163-21

Cited by 22 publications

(125 citation statements)

References 108 publications

(120 reference statements)

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“…This suggests that both E2WT and E2S23A can bind to the labeled plasmid and retain it in the nucleus. Both E2WT and E2S23A activate transcription to an identical level in a 3-day transcription assay in U2OS cells (42). However, it was noticeable that in U2OS-E2-WT mitotic cells, but not in U2OS-E2-S23A mitotic cells, the fluorescent ptk6E2-luc was retained on the mitotic chromatin (mitotic cells indicated with white arrows).…”

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 98%

“…Previously we demonstrated co-localization of E2 and TopBP1 on mitotic chromatin, and shown that E2 can recruit TopBP1 to mitotic chromatin (41,42). There were no assays available to determine whether E2 recruits plasmids onto mitotic chromatin; we therefore developed an assay.…”

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

“…In these experiments, we could not co-stain for E2 protein as the permeabilization disrupted the interaction of the fluorescent plasmids with the chromatin. These cell lines stably express E2, and most cells retain E2 protein expression (42).…”

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

“…At day 9, all cells were harvested for luciferase assays. The ability of E2-WT and E2-S23A to activate transcription over a day three period is identical, this was therefore set as the reference point 1, to which the activity at day 6 and 9 is presented as relative (42). The upper panel of Figure 2A demonstrates that, in U2OS-E2-WT cells, around 85% of luciferase activity is retained between days 3 and 6, while in U2OS-E2-S23A cells retention is around 15% (note the log scale).…”

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

“…Furthermore, TopBP1 regulates the ability of E2 to interact with interphase chromatin, and co-localizes with E2 on mitotic chromatin (41). Recently, we demonstrated that phosphorylation of E2 on serine 23 promotes a direct interaction between these two proteins in vitro and in vivo, and that E2 recruits TopBP1 onto mitotic chromatin (42). Our work showed that mutation of serine 23 to alanine (S23A) resulted in a compromised interaction of E2 with mitotic chromatin, and that E2-TopBP1 interaction is essential for the HPV16 life cycle (42).…”

Section: Introductionmentioning

confidence: 99%

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Interaction with TopBP1 mediates human papillomavirus 16 E2 plasmid segregation/retention function and stability during the viral life cycle

Prabhakar

James

Das

et al. 2022

Preprint

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Human papillomavirus 16 (HPV16) E2 is a DNA binding protein that regulates transcription, replication and potentially, segregation of the HPV16 genome during the viral life cycle. In the segregation model, E2 simultaneously binds to viral and host chromatin, acting as a bridge to ensure that viral genomes reside in daughter nuclei following cell division. The host chromatin receptor for E2 mediating this function is unknown. Recently, we demonstrated that CK2 phosphorylation of E2 on serine 23 (S23) is required for interaction with TopBP1, and that this interaction promotes E2 and TopBP1 recruitment to mitotic chromatin. Here, we demonstrate that in U2OS and N/Tert-1 cells expressing wild type E2 and a non-TopBP1 binding mutant (S23A, serine 23 mutated to alanine), interaction with TopBP1 is essential for E2 recruitment of plasmids to mitotic chromatin. Using a novel quantitative segregation assay, we demonstrate that interaction with TopBP1 is required for E2 plasmid segregation. The interaction of E2 with TopBP1 promotes increased levels of E2 protein during mitosis in U2OS and N/Tert-1 cells, as well as in human foreskin keratinocytes immortalized by the HPV16 genome. Additionally, interaction with TopBP1 is required for expression of the E2 protein during the viral life cycle. Overall, our results demonstrate that E2 has plasmid segregation activity, and that the E2-TopBP1 interaction is essential for this E2 function and for E2 expression during the viral life cycle.

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Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 98%

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

Section: Development Of Functional Assays For Investigating E2 Segregation Functionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interaction with TopBP1 mediates human papillomavirus 16 E2 plasmid segregation/retention function and stability during the viral life cycle

Prabhakar

James

Das

et al. 2022

Preprint

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Human papillomavirus associated cervical lesion: pathogenesis and therapeutic interventions

Ye,

Zheng,

et al. 2023

MedComm

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Human papillomavirus (HPV) is the most prevalent sexually transmitted virus globally. Persistent high‐risk HPV infection can result in cervical precancerous lesions and cervical cancer, with 70% of cervical cancer cases associated with high‐risk types HPV16 and 18. HPV infection imposes a significant financial and psychological burden. Therefore, studying methods to eradicate HPV infection and halt the progression of precancerous lesions remains crucial. This review comprehensively explores the mechanisms underlying HPV‐related cervical lesions, including the viral life cycle, immune factors, epithelial cell malignant transformation, and host and environmental contributing factors. Additionally, we provide a comprehensive overview of treatment methods for HPV‐related cervical precancerous lesions and cervical cancer. Our focus is on immunotherapy, encompassing HPV therapeutic vaccines, immune checkpoint inhibitors, and advanced adoptive T cell therapy. Furthermore, we summarize the commonly employed drugs and other nonsurgical treatments currently utilized in clinical practice for managing HPV infection and associated cervical lesions. Gene editing technology is currently undergoing clinical research and, although not yet employed officially in clinical treatment of cervical lesions, numerous preclinical studies have substantiated its efficacy. Therefore, it holds promise as a precise treatment strategy for HPV‐related cervical lesions.

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Protein kinase CK2 – diverse roles in cancer cell biology and therapeutic promise

et al. 2022

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The association of protein kinase CK2 (formerly casein kinase II or 2) with cell growth and proliferation in cells was apparent at early stages of its investigation. A cancer-specific role for CK2 remained unclear until it was determined that CK2 was also a potent suppressor of cell death (apoptosis); the latter characteristic differentiated its function in normal versus malignant cells because dysregulation of both cell growth and cell death is a universal feature of cancer cells. Over time, it became evident that CK2 exerts its influence on a diverse range of cell functions in normal as well as in transformed cells. As such, CK2 and its substrates are localized in various compartments of the cell. The dysregulation of CK2 is documented in a wide range of malignancies; notably, by increased CK2 protein and activity levels with relatively moderate change in its RNA abundance. High levels of CK2 are associated with poor prognosis in multiple cancer types, and CK2 is a target for active research and testing for cancer therapy. Aspects of CK2 cellular roles and targeting in cancer are discussed in the present review, with focus on nuclear and mitochondrial functions and prostate, breast and head and neck malignancies.

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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

Cited by 22 publications

References 108 publications

Interaction with TopBP1 mediates human papillomavirus 16 E2 plasmid segregation/retention function and stability during the viral life cycle

Interaction with TopBP1 mediates human papillomavirus 16 E2 plasmid segregation/retention function and stability during the viral life cycle

Human papillomavirus associated cervical lesion: pathogenesis and therapeutic interventions

Protein kinase CK2 – diverse roles in cancer cell biology and therapeutic promise

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