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J Gen Virol 87 (2006), 789-793; DOI 10.1099/vir.0.81640-0

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© 2006 Society for General Microbiology

Short Communication

Nuclear localization of the Epstein–Barr virus EBNA3B protein

Anita Burgess1, Marion Buck1, Kenia Krauer1 and Tom Sculley1,2

1 Queensland Institute of Medical Research, 300 Herston Road, Brisbane, Queensland 4029, Australia
2 Griffith Medical Research Centre, 300 Herston Road, Brisbane, Queensland 4029, Australia

Correspondence
Anita Burgess
anitaB{at}qimr.edu.au

The Epstein–Barr virus nuclear antigen (EBNA) 3B is a hydrophilic, proline-rich, charged protein that is thought to be involved in transcriptional regulation and is targeted exclusively to the cell nucleus, where it localizes to discrete subnuclear granules. Co-localization studies utilizing a fusion protein between enhanced green fluorescent protein (EGFP) and EBNA3B with FLAG-tagged EBNA3A and EBNA3C proteins demonstrated that EBNA3B co-localized with both EBNA3A and EBNA3C in the nuclei of cells when overexpressed. Computer analyses identified four potential nuclear-localization signals (NLSs) in the EBNA3B amino acid sequence. By utilizing fusion proteins with EGFP, deletion constructs of EBNA3B and site-directed mutagenesis, three of the four NLSs (aa 160–166, 430–434 and 867–873) were shown to be functional in truncated forms of EBNA3B, whilst an additional NLS (aa 243–246) was identified within the N-terminal region of EBNA3B. Only two of the NLSs were found to be functional in the context of the full-length EBNA3B protein.






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