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Nucleic Acids Research, 1988, Vol. 16, No. 8 3415-3435
© 1988

Articles

Mutational analysis of Epstein-Barr virus nuclear antigen 1 (EBNA 1)

Maryellen Polvino-Bodnar, Janette Kiso and Priscilla A. Schaffer*

Laboratory of Tumor Virus Genetics, Dana-Farber Cancer Institute and Department of Microbiology and Molecular Genetics Harvard Medical School, Boston, MA 02115, USA

*To whom correspondence should be addressed

Received October 29, 1987. Revised March 10, 1988. Accepted March 10, 1988.

We have constructed a set of nonsense mutants in the EBNA 1 gene of Epstein-Barr virus by inserting a synthetic oligonucleotide, which has transnational termination codons in all three reading frames, at various positions in a cloned copy of the EBNA 1 gene. The EBNA 1 proteins encoded by these mutants and three deletion mutants were analyzed using several functional assays. It was determined that there are two seperable phosphorylation domains in the carboxy half of the molecule. The carboxy half of the molecule was also found to contain a region between the unique Sac I and Sac II sites that is required for transactivation of the EBNA 1-specific enhancer element found within ori P. The mutants also served to identify a 248 bp region that affects the pattern of intranuclear localization of the protein. Correlations between the functional domains established by these studies and other properties of EBNA 1 are discussed.


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