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J. Virol., Apr 1996, 2562-2568, Vol 70, No. 4
L Szekely, K Pokrovskaja, WQ Jiang, H de The, N Ringertz and G Klein
EBNA-5 is one of the Epstein-Barr virus (EBV)-encoded nuclear proteins
required for immortalization of human B lymphocytes. In the nuclei of
EBV-transformed lymphoblastoid cell lines EBNA-5 is preferentially
targetted to distinct nuclear foci. Previously we have shown (W.Q. Jiang,
L. Szekely, V. Wendel-Hansen, N. Ringertz, G. Klein, and A. Rosen, Exp.
Cell Res. 197:314-318, 1991) that the same foci also contained the
retinoblastoma (Rb) protein. Using a similar double immunofluorescence
technique, we now show that these foci colocalize with nuclear bodies
positive for PML, the promyelocytic leukemia- associated protein.
Artificial spreading of the chromatin by exposure to the forces of fluid
surface tension disrupts this colocalization gradually, suggesting that the
bodies consist of at least two subcomponents. Heat shock or metabolic
stress induced by high cell density leads to the release of EBNA-5 from the
PML-positive nuclear bodies and induces it to translocate to the nucleoli.
In addition to their presence in nuclear bodies, both proteins are
occasionally present in nuclear aggregates and doughnut-like structures in
which PML is concentrated in an outer shell. Nuclear bodies with prominent
PML staining are seen in resting B lymphocytes. This staining pattern does
not change upon EBV infection. In freshly infected cells EBNA-5 antigens
are first distributed throughout the nucleoplasm. After a few days
intensely staining foci develop. These foci coincide with PML- positive
nuclear bodies. At a later stage and in established lymphoblastoid cell
lines EBNA-5 is almost exclusively present in the PML-positive nuclear
foci. The colocalization is restricted to EBV- infected human lymphoblasts.
The data presented indicate that the distinct EBNA-5 foci are not newly
formed structures but the result of translocation of the viral protein to a
specialized domain present already in the nuclei of uninfected cells.
Copyright © 1996, American Society for Microbiology
The Epstein-Barr virus-encoded nuclear antigen EBNA-5 accumulates in PML-containing bodies
Microbiology and Tumor Biology Center, Karolinska Institute, Stockholm, Sweden.
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