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Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):3221-6. doi: 10.1073/pnas.1601773113. Epub 2016 Mar 7.

EBV noncoding RNA EBER2 interacts with host RNA-binding proteins to regulate viral gene expression.

Author information

1
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06536;
2
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06536; Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06536 joan.steitz@yale.edu.

Abstract

Epstein-Barr virus (EBV) produces a highly abundant noncoding RNA called EBV-encoded RNA 2 (EBER2) that interacts indirectly with the host transcription factor paired box protein 5 (PAX5) to regulate viral latent membrane protein 1/2 (LMP1/2) gene expression as well as EBV lytic replication. To identify intermediary proteins, we isolated EBER2-PAX5-containing complexes and analyzed the protein components by mass spectrometry. The top candidates include three host proteins splicing factor proline and glutamine rich (SFPQ), non-POU domain-containing octamer-binding protein (NONO), and RNA binding motif protein 14 (RBM14), all reported to be components of nuclear bodies called paraspeckles. In vivo RNA-protein crosslinking indicates that SFPQ and RBM14 contact EBER2 directly. Binding studies using recombinant proteins demonstrate that SFPQ and NONO associate with PAX5, potentially bridging its interaction with EBER2. Similar to EBER2 or PAX5 depletion, knockdown of any of the three host RNA-binding proteins results in the up-regulation of viral LMP2A mRNA levels, supporting a physiologically relevant interaction of these newly identified factors with EBER2 and PAX5. Identification of these EBER2-interacting proteins enables the search for cellular noncoding RNAs that regulate host gene expression in a manner similar to EBER2.

KEYWORDS:

Epstein–Barr virus; RNA binding protein; noncoding RNA; paraspeckle

PMID:
26951683
PMCID:
PMC4812724
DOI:
10.1073/pnas.1601773113
[Indexed for MEDLINE]
Free PMC Article

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