Abstract
EBNA-3 (also called EBNA-3A) is one of the EBV encoded nuclear antigens that are necessary for B-cell transformation. EBNA-3 is known to target RBPs, nuclear proteins that also interacts with EBNA-2, EBNA-4 and EBNA-6. In order to identify additional EBNA-3 targets, an EBV-transformed human lymphocyte cDNA library was screened in the yeast two-hybrid system with N-terminus truncated EBNA-3 that cannot interact with RBP-Jkappa. A clone, encoding Xap-2 protein, a cellular partner of Hepatitis B virus X-antigen was isolated. This protein is also known as the p38 subunit of the aryl hydrocarbon receptor complex (ARA9). The specific binding to EBNA-3 was confirmed by showing that the GST-Xap-2 precipitated EBNA-3 from CV1 cells that were infected with recombinant vaccinia virus expressing EBNA-3. Deletion of the C-terminus of Xap-2 eliminated the binding. Fusion with green fluorescent protein showed that Xap-2 is preferentially cytoplasmic but translocates to the nucleus upon expression of EBNA-3.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Amino Acid Sequence
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Animals
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Base Sequence
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Cell Line
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Cell Line, Transformed
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DNA, Complementary
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Epstein-Barr Virus Nuclear Antigens / genetics
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Epstein-Barr Virus Nuclear Antigens / metabolism*
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Hepatitis B Antigens / metabolism*
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Herpesvirus 4, Human / genetics
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Herpesvirus 4, Human / metabolism*
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Humans
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Intracellular Signaling Peptides and Proteins
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Mice
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Molecular Sequence Data
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Proteins / genetics
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Proteins / metabolism*
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Receptors, Aryl Hydrocarbon / genetics
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Receptors, Aryl Hydrocarbon / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism*
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Subcellular Fractions
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Trans-Activators / metabolism*
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Two-Hybrid System Techniques
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Viral Regulatory and Accessory Proteins
Substances
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Carrier Proteins
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DNA, Complementary
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Epstein-Barr Virus Nuclear Antigens
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Hepatitis B Antigens
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Intracellular Signaling Peptides and Proteins
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Proteins
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Receptors, Aryl Hydrocarbon
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Recombinant Fusion Proteins
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Trans-Activators
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Viral Regulatory and Accessory Proteins
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aryl hydrocarbon receptor-interacting protein
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hepatitis B virus X protein