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Published 8 May 2006. doi:10.1083/jcb.200601026
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 173, Number 3, 319-325
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Epstein-Barr virus noncoding RNAs are confined to the nucleus, whereas their partner, the human La protein, undergoes nucleocytoplasmic shuttling



Victor Fok1,2, Kyle Friend1,2, and Joan A. Steitz1,2

1 Department of Molecular Biophysics and Biochemistry and 2 Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06536

Correspondence to Joan A. Steitz: joan.steitz{at}yale.edu

The Epstein-Barr virus (EBV) noncoding RNAs, EBV-encoded RNA 1 (EBER1) and EBER2, are the most abundant viral transcripts in all types of latently infected human B cells, but their function remains unknown. We carried out heterokaryon assays using cells that endogenously produce EBERs to address their trafficking, as well as that of the La protein, because EBERs are quantitatively bound by La in vivo. Both in this assay and in oocyte microinjection assays, EBERs are confined to the nucleus, suggesting that their contribution to viral latency is purely nuclear. EBER1 does not bind exportin 5; therefore, it is unlikely to act by interfering with microRNA biogenesis. In contrast, La, which is a nuclear phosphoprotein, undergoes nucleocytoplasmic shuttling independent of the nuclear export protein Crm1. To ensure that small RNA shuttling can be detected in cells that are negative for EBER shuttling, we demonstrate the shuttling of U1 small nuclear RNA.

Abbreviations used in this paper: DIG, digoxigenin; EBER, EBV-encoded RNA; EBV, Epstein-Barr virus; Exp5, exportin 5; hnRNP, heterogeneous nuclear ribonucleoprotein; HEK, human embryonic kidney; LMB, leptomycin B; PKR, protein kinase R; snRNA, small nuclear RNA.


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