Carcinoma-risk variant of EBNA1 deregulates Epstein-Barr Virus episomal latency

Dheekollu, Jayaraju; Malecka, Kimberly A.; Wiedmer, Andreas; Delecluse, Henri Jacques; Chiang, Alan Kwok Shing; Altieri, Dario C.; Messick, Troy E.; Lieberman, Paul M.

doi:10.18632/oncotarget.14540

Cited by 51 publications

(61 citation statements)

References 54 publications

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“…Recently, our laboratory found that an NPC variant of EBNA1 containing 8 amino acid polymorphisms different from the prototypical B95-8 strain, including a T585I substitution, was defective in replication and maintenance of the viral episome (41). The EBNA1 variant was also found to be deficient in suppressing lytic cycle gene transcription and lytic DNA replication (41). We note that the Mutu strain isolated from Burkitt's lymphoma contains both T585P and L582F among 8 additional polymorphisms.…”

Section: Discussionmentioning

confidence: 81%

“…Genetic analyses of various EBV strains indicate that T585 is among the EBNA1 polymorphisms found frequently in both NPC tumors and Burkitt's lymphoma (35)(36)(37)(38)(39)(40)(41). Recently, our laboratory found that an NPC variant of EBNA1 containing 8 amino acid polymorphisms different from the prototypical B95-8 strain, including a T585I substitution, was defective in replication and maintenance of the viral episome (41). The EBNA1 variant was also found to be deficient in suppressing lytic cycle gene transcription and lytic DNA replication (41).…”

Section: Discussionmentioning

confidence: 99%

“…Cells were cultured in an incubator set at 37°C with 5% CO 2 . The mammalian expression vector for Flag-EBNA1 or hemagglutinin (HA)-EBNA1 contained B95-8 EBNA1 lacking the GA repeats (aa 101 to 324) under the control of a CMV-3ϫFlag or a CMV-HA promoter in a plasmid derived from pREP10 (Clontech) containing OriP, green fluorescent protein (GFP), and hygromycin resistance (41).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid maintenance assays were described previously (41,50). Briefly, HeLa or 293T cells were seeded into 6-well plates at concentrations of 0.75 ϫ 10 5 to 1.5 ϫ 10 5 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid DNA replication assays were described previously (41,51). Briefly, HeLa and 293T cells (ϳ1 ϫ 10 6 cells) were plated into 10-cm dishes.…”

Section: Methodsmentioning

confidence: 99%

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Structural and Functional Basis for an EBNA1 Hexameric Ring in Epstein-Barr Virus Episome Maintenance

Deakyne

Malecka

Messick

et al. 2017

J Virol

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Epstein-Barr virus (EBV) establishes a stable latent infection that can persist for the life of the host. EBNA1 is required for the replication, maintenance, and segregation of the latent episome, but the structural features of EBNA1 that confer each of these functions are not completely understood. Here, we have solved the X-ray crystal structure of an EBNA1 DNA-binding domain (DBD) and discovered a novel hexameric ring oligomeric form. The oligomeric interface pivoted around residue T585 as a joint that links and stabilizes higher-order EBNA1 complexes. Substitution mutations around the interface destabilized higher-order complex formation and altered the cooperative DNA-binding properties of EBNA1. Mutations had both positive and negative effects on EBNA1-dependent DNA replication and episome maintenance with OriP. We found that one naturally occurring polymorphism in the oligomer interface (T585P) had greater cooperative DNA binding in vitro, minor defects in DNA replication, and pronounced defects in episome maintenance. The T585P mutant was compromised for binding to OriP in vivo as well as for assembling the origin recognition complex subunit 2 (ORC2) and trimethylated histone 3 lysine 4 (H3K4me3) at OriP. The T585P mutant was also compromised for forming stable subnuclear foci in living cells. These findings reveal a novel oligomeric structure of EBNA1 with an interface subject to naturally occurring polymorphisms that modulate EBNA1 functional properties. We propose that EBNA1 dimers can assemble into higher-order oligomeric structures important for diverse functions of EBNA1.IMPORTANCE Epstein-Barr virus is a human gammaherpesvirus that is causally associated with various cancers. Carcinogenic properties are linked to the ability of the virus to persist in the latent form for the lifetime of the host. EBNA1 is a sequencespecific DNA-binding protein that is consistently expressed in EBV tumors and is the only viral protein required to maintain the viral episome during latency. The structural and biochemical mechanisms by which EBNA1 allows the long-term persistence of the EBV genome are currently unclear. Here, we have solved the crystal structure of an EBNA1 hexameric ring and characterized key residues in the interface required for higher-order complex formation and long-term plasmid maintenance.KEYWORDS DNA-binding domain, EBNA1, Epstein-Barr virus, OriP, cooperative binding, hexameric ring, oligomers, episome maintenance, viral latency E pstein-Barr virus (EBV) is a ubiquitous human herpesvirus that establishes long-term latent infection in more than 90% of the human population (1-6). Latent infection by EBV is estimated to be responsible for over 200,000 cancers worldwide, including subtypes of Burkitt's lymphoma, diffuse large B-cell lymphoma, primary central nervous system lymphoma, and Hodgkin's lymphoma as well as gastric carcinoma and nasopharyngeal carcinoma (NPC) (1-4). Epstein-Barr virus nuclear antigen 1 (EBNA1) is the only viral protein consistently expressed in all EBV-associate...

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Plasmid maintenance assays were described previously (41,50). Briefly, HeLa or 293T cells were seeded into 6-well plates at concentrations of 0.75 ϫ 10 5 to 1.5 ϫ 10 5 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid DNA replication assays were described previously (41,51). Briefly, HeLa and 293T cells (ϳ1 ϫ 10 6 cells) were plated into 10-cm dishes.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Structural and Functional Basis for an EBNA1 Hexameric Ring in Epstein-Barr Virus Episome Maintenance

Deakyne

Malecka

Messick

et al. 2017

J Virol

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A novel Epstein‐Barr virus subtype associated with nasopharyngeal carcinoma found in South China

2020

Cancer Communications

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Advances in pathogenesis and precision medicine for nasopharyngeal carcinoma

Zhu

Zhao

et al. 2021

MedComm

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Nasopharyngeal carcinoma (NPC) is a squamous carcinoma with apparent geographical and racial distribution, mostly prevalent in East and Southeast Asia, particularly concentrated in southern China. The epidemiological trend over the past decades has suggested a substantial reduction in the incidence rate and mortality rate due to NPC. These results may reflect changes in lifestyle and environment, and more importantly, a deeper comprehension of the pathogenic mechanism of NPC, leading to much progress in the preventing, screening, and treating for this cancer. Herein, we present the recent advances on the key signal pathways involved in pathogenesis of NPC, the mechanism of Epstein‐Barr virus (EBV) entry into the cell, and the progress of EBV vaccine and screening biomarkers. We will also discuss in depth the development of various therapeutic approaches including radiotherapy, chemotherapy, surgery, targeted therapy, and immunotherapy. These research advancements have led to a new era of precision medicine in NPC.

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Carcinoma-risk variant of EBNA1 deregulates Epstein-Barr Virus episomal latency

Cited by 51 publications

References 54 publications

Structural and Functional Basis for an EBNA1 Hexameric Ring in Epstein-Barr Virus Episome Maintenance

Structural and Functional Basis for an EBNA1 Hexameric Ring in Epstein-Barr Virus Episome Maintenance

A novel Epstein‐Barr virus subtype associated with nasopharyngeal carcinoma found in South China

Advances in pathogenesis and precision medicine for nasopharyngeal carcinoma

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