Structural Basis for Cooperative Binding of EBNA1 to the Epstein-Barr Virus Dyad Symmetry Minimal Origin of Replication

Malecka, Kimberly A.; Dheekollu, Jayaraju; Deakyne, Julianna S.; Wiedmer, Andreas; Ramirez, Ursula D.; Lieberman, Paul M.; Messick, Troy E.

doi:10.1128/jvi.00487-19

Title: Structural Basis for Cooperative Binding of EBNA1 to the Epstein-Barr Virus Dyad Symmetry Minimal Origin of Replication

Journal Article · Mon Sep 30 00:00:00 EDT 2019 · Journal of Virology

DOI:https://doi.org/10.1128/jvi.00487-19· OSTI ID:1570743

Malecka, Kimberly A. ^[1]; Dheekollu, Jayaraju ^[1]; Deakyne, Julianna S. ^[1]; Wiedmer, Andreas ^[1]; Ramirez, Ursula D. ^[2]; Lieberman, Paul M. ^[1]; Messick, Troy E. ^[1]

The Wistar Inst., Philadelphia, PA (United States)
Vironika, LLC, Wynnewood, PA (United States)

Epstein-Barr virus is associated with several human malignancies, including nasopharyngeal carcinoma, gastric cancer, and lymphoma. Latently infected cells carry a circularized EBV episome where the origin of replication (oriP) is comprised of two elements: the family of repeats (FR) and dyad symmetry (DS). The viral protein Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) binds to FR and DS to promote EBV episome maintenance and DNA replication during latent infection in proliferating cells. EBNA1 binding to the DS constitutes a minimal origin of DNA replication. Here we report the crystal structure of two EBNA1 DNA-binding domain dimers bound to a DS half-site. This structure shows that the DNA is smoothly bent, allowing for stabilizing interactions between the dimers. The dimer-dimer interface requires an intricate hydrogen bonding network involving residues R491 and D581. When this interface is disrupted, we note loss of stable dimer-dimer complex formation on the DNA, compromised oriP-containing plasmid replication in cells, and impaired recruitment of the MCM3 complex to the oriP. Surface conservation analysis reveals that these residues are part of a larger conserved surface that may be critical for recruitment of replication machinery to the oriP. Our results reveal a new region of EBNA1 critical for its activity and one that may be exploited by targeted small molecules to treat EBV-associated disease.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH)

Grant/Contract Number:: RO1 CA093606; RO1 DE017336; P30 CA10815; WT096496; T32 CA09171; WO2015073864; WO2016183534

OSTI ID:: 1570743

Journal Information:: Journal of Virology, Vol. 93, Issue 20; ISSN 0022-538X

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 8 works

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59 BASIC BIOLOGICAL SCIENCES
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Title: Structural Basis for Cooperative Binding of EBNA1 to the Epstein-Barr Virus Dyad Symmetry Minimal Origin of Replication

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