@article{mbs:/content/journal/jgv/10.1099/vir.0.19549-0, author = "Krauer, Kenia and Buck, Marion and Flanagan, James and Belzer, Deanna and Sculley, Tom", title = "Identification of the nuclear localization signals within the Epstein–Barr virus EBNA-6 protein", journal= "Journal of General Virology", year = "2004", volume = "85", number = "1", pages = "165-172", doi = "https://doi.org/10.1099/vir.0.19549-0", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.19549-0", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = "Epstein–Barr virus nuclear antigen (EBNA)-6 is essential for EBV-induced immortalization of primary human B-lymphocytes in vitro. Previous studies have shown that EBNA-6 acts as a transcriptional regulator of viral and cellular genes; however at present, few functional domains of the 140 kDa EBNA-6 protein have been completely characterized. There are five computer-predicted nuclear localization signals (NLS), four monopartite and one bipartite, present in the EBNA-6 amino acid sequence. To identify which of these NLS are functional, fusion proteins between green fluorescent protein and deletion constructs of EBNA-6 were expressed in HeLa cells. Each of the constructs containing at least one of the NLS was targeted to the nucleus of cells whereas a construct lacking all of the NLS was cytoplasmic. Site-directed mutation of these NLS demonstrated that only three of the NLS were functional, one at the N-terminal end (aa 72–80), one in the middle (aa 412–418) and one at the C-terminal end (aa 939–945) of the EBNA-6 protein.", }