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J Virol. 2019 Jun 28;93(14). pii: e00724-19. doi: 10.1128/JVI.00724-19. Print 2019 Jul 15.

A Noncanonical Basic Motif of Epstein-Barr Virus ZEBRA Protein Facilitates Recognition of Methylated DNA, High-Affinity DNA Binding, and Lytic Activation.

Author information

1
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA.
2
Department of Pediatrics, Yale University, New Haven, Connecticut, USA.
3
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA george.miller@yale.edu yong.xiong@yale.edu.
4
Department of Epidemiology and Public Health, Yale University, New Haven, Connecticut, USA.
#
Contributed equally

Abstract

The pathogenesis of Epstein-Barr virus (EBV) infection, including development of lymphomas and carcinomas, is dependent on the ability of the virus to transit from latency to the lytic phase. This conversion, and ultimately disease development, depends on the molecular switch protein, ZEBRA, a viral bZIP transcription factor that initiates transcription from promoters of viral lytic genes. By binding to the origin of viral replication, ZEBRA is also an essential replication protein. Here, we identified a novel DNA-binding motif of ZEBRA, N terminal to the canonical bZIP domain. This RRTRK motif is important for high-affinity binding to DNA and is essential for recognizing the methylation state of viral promoters. Mutations in this motif lead to deficiencies in DNA binding, recognition of DNA methylation, lytic cycle DNA replication, and viral late gene expression. This work advances our understanding of ZEBRA-dependent activation of the viral lytic cascade.IMPORTANCE The binding of ZEBRA to methylated and unmethylated viral DNA triggers activation of the EBV lytic cycle, leading to viral replication and, in some patients, cancer development. Our work thoroughly examines how ZEBRA uses a previously unrecognized basic motif to bind nonmethylated and methylated DNA targets, leading to viral lytic activation. Our findings show that two different positively charged motifs, including the canonical BZIP domain and a newly identified RRTRK motif, contribute to the mechanism of DNA recognition by a viral AP-1 protein. This work contributes to the assessment of ZEBRA as a potential therapeutic target for antiviral and oncolytic treatments.

KEYWORDS:

AP-1-like protein; BZLF1; DNA methylation; DNA-binding domain; Epstein-Barr virus; ZEBRA; bZIP protein; lytic activation; noncanonical DNA-binding domain; transcription factors

PMID:
31068430
PMCID:
PMC6600195
[Available on 2019-12-28]
DOI:
10.1128/JVI.00724-19

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