Modelling the structure of full-length Epstein–Barr virus nuclear antigen 1

Hussain, Mushtaq; Gatherer, Derek; Wilson, Joanna B.

doi:10.1007/s11262-014-1101-9

Cited by 14 publications

(18 citation statements)

References 76 publications

(72 reference statements)

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“…6, top). A recent study used molecular modelling to assess the overall structure of full-length EBNA1 bound to DNA further addressing the 3D-folding, and DNAprotein interactions largely confirmed the above findings (Hussain et al 2014). …”

Section: Ebna1mentioning

confidence: 58%

Epstein-Barr Virus-Specific Humoral Immune Responses in Health and Disease

Middeldorp

2015

Epstein Barr Virus Volume 2

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Epstein-Barr virus (EBV) is widely distributed in the world and associated with a still increasing number of acute, chronic, malignant and autoimmune disease syndromes. Humoral immune responses to EBV have been studied for diagnostic, pathogenic and protective (vaccine) purposes. These studies use a range of methodologies, from cell-based immunofluorescence testing to antibody-diversity analysis using immunoblot and epitope analysis using recombinant or synthetic peptide-scanning. First, the individual EBV antigen complexes (VCA , MA, EA(D), EA(R) and EBNA) are defined at cellular and molecular levels, providing a historic overview. The characteristic antibody responses to these complexes in health and disease are described, and differences are highlighted by clinical examples. Options for EBV vaccination are briefly addressed. For a selected number of immunodominant proteins, in particular EBNA1, the interaction with human antibodies is further detailed at the epitope level, revealing interesting insights for structure, function and immunological aspects, not considered previously. Humoral immune responses against EBV-encoded tumour antigens LMP1, LMP2 and BARF1 are addressed, which provide novel options for targeted immunotherapy. Finally, some considerations on EBV-linked autoimmune diseases are given, and mechanisms of antigen mimicry are briefly discussed. Further analysis of humoral immune responses against EBV in health and disease in carefully selected patient cohorts will open new options for understanding pathogenesis of individual EBV-linked diseases and developing targeted diagnostic and therapeutic approaches.

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Section: Ebna1mentioning

confidence: 58%

Epstein-Barr Virus-Specific Humoral Immune Responses in Health and Disease

Middeldorp

2015

Epstein Barr Virus Volume 2

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“…The dimer-dimer interaction at the DS is essential for the initiation of DNA replication, and this cooperativity is mediated, in part, through predicted interactions in the DNAbinding domain (8). Cooperative interactions that enable DNA looping between distal EBNA1 sites are known to be mediated through the amino-terminal linking domains of EBNA1, which are capable of forming homotypic Zn-hook interactions (22,23). Cooperative interactions between EBNA1 DNA-binding domains at the FR are not yet known.…”

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confidence: 99%

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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“…EBNA1 is a sequence specific DNA binding protein, which acts as a homodimer to cradle the bound viral genome using the C-terminal domain (residues 459 to 607) and thereby carries and attaches the viral genome to the cellular chromatin via the N-terminal and central domains. The crystal structure of the C-terminal domain has been resolved bound to DNA [ 38 , 39 ] and the full protein modelled in silico [ 40 ], to reveal that this C-terminal DNA binding domain connects to the remainder of the protein by a string of residues that runs through the major groove of the DNA. In addition, higher order structures have been observed, specifically trimmers of dimers [ 41 ].…”

Section: Introductionmentioning

confidence: 99%

“…Several residues in the C-terminal DNA binding domain have been shown to be important for interactions in the dimer [ 42 ]. Additionally, contact residues between monomers in the central region of the protein have been identified in the modelled dimer [ 40 ]. Within the C-terminal domain is a proline-rich stretch which forms a protruding loop (seen in both the crystal structure and the in silico model), which has been proposed to anchor one monomer to the other in the dimer, with the acidic C-terminal tail of one monomer, part wrapping around the proline loop of the other [ 40 ].…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, contact residues between monomers in the central region of the protein have been identified in the modelled dimer [ 40 ]. Within the C-terminal domain is a proline-rich stretch which forms a protruding loop (seen in both the crystal structure and the in silico model), which has been proposed to anchor one monomer to the other in the dimer, with the acidic C-terminal tail of one monomer, part wrapping around the proline loop of the other [ 40 ]. The EBNA1 dimers link together when bound to repeats of the DNA recognition sequence, as found in the viral origin of plasmid replication (oriP).…”

Section: Introductionmentioning

confidence: 99%

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EBNA1: Oncogenic Activity, Immune Evasion and Biochemical Functions Provide Targets for Novel Therapeutic Strategies against Epstein-Barr Virus- Associated Cancers

Wilson

Manet

Gruffat

et al. 2018

Cancers

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The presence of the Epstein-Barr virus (EBV)-encoded nuclear antigen-1 (EBNA1) protein in all EBV-carrying tumours constitutes a marker that distinguishes the virus-associated cancer cells from normal cells and thereby offers opportunities for targeted therapeutic intervention. EBNA1 is essential for viral genome maintenance and also for controlling viral gene expression and without EBNA1, the virus cannot persist. EBNA1 itself has been linked to cell transformation but the underlying mechanism of its oncogenic activity has been unclear. However, recent data are starting to shed light on its growth-promoting pathways, suggesting that targeting EBNA1 can have a direct growth suppressing effect. In order to carry out its tasks, EBNA1 interacts with cellular factors and these interactions are potential therapeutic targets, where the aim would be to cripple the virus and thereby rid the tumour cells of any oncogenic activity related to the virus. Another strategy to target EBNA1 is to interfere with its expression. Controlling the rate of EBNA1 synthesis is critical for the virus to maintain a sufficient level to support viral functions, while at the same time, restricting expression is equally important to prevent the immune system from detecting and destroying EBNA1-positive cells. To achieve this balance EBNA1 has evolved a unique repeat sequence of glycines and alanines that controls its own rate of mRNA translation. As the underlying molecular mechanisms for how this repeat suppresses its own rate of synthesis in cis are starting to be better understood, new therapeutic strategies are emerging that aim to modulate the translation of the EBNA1 mRNA. If translation is induced, it could increase the amount of EBNA1-derived antigenic peptides that are presented to the major histocompatibility (MHC) class I pathway and thus, make EBV-carrying cancers better targets for the immune system. If translation is further suppressed, this would provide another means to cripple the virus.

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Modelling the structure of full-length Epstein–Barr virus nuclear antigen 1

Cited by 14 publications

References 76 publications

Epstein-Barr Virus-Specific Humoral Immune Responses in Health and Disease

Epstein-Barr Virus-Specific Humoral Immune Responses in Health and Disease

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

EBNA1: Oncogenic Activity, Immune Evasion and Biochemical Functions Provide Targets for Novel Therapeutic Strategies against Epstein-Barr Virus- Associated Cancers

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