P32/TAP, a cellular protein that interacts with EBNA-1 of Epstein-Barr virus

Yilong Wang, Jon E. Finan, Jaap M. Middeldorp, S. Diane Hayward

Research output: Contribution to journalArticle

Abstract

The Epstein-Barr virus (EBV) EBNA-1 protein has a central role in the maintenance of a latent EBV infection and is the only virus-encoded protein expressed in all EBV-associated tumors. EBNA-1 is required for replication of the episomal form of the latent viral genome and transactivates the latency C and LMP-1 promoters. The mechanisms by which EBNA-1 performs these functions are not known. Here we describe the cloning, expression, and characterization of a cellular protein, P32/TAP, which strongly interacts with EBNA-1. We show that P32/TAP is expressed at high levels in Raji cells and is synthesized as a proprotein of 282 amino acids (aa) that is posttranslationally processed by a two-step cleavage process to yield a mature protein of 209 aa. It has been previously reported that P32/TAP is expressed on the cell surface. Our transient expression assays detected full-length P32/TAP (1-282 aa) in the cytoplasm while mature P32/TAP protein localized to the nucleus. Three lines of evidence support P32/TAP interaction with EBNA-1. First, in the yeast two-hybrid system we mapped two interactive N-terminal regions of EBNA-1, aa 40-60 and aa 325-376, each of which contains arginine-glycine repeats. These regions interact with the C-terminal half of P32/TAP. Second, the full-length cytoplasmic P32/TAP protein can translocate nuclear EBNA-1 into the cytoplasm. Third, P32/TAP co-immunoprecipitated with EBNA-1. We have confirmed that a Gal4 fusion protein containing the C-terminal region of P32/TAP (aa 244-282) transactivates expression from a reporter containing upstream Gal4-binding sites. Deletion of the P32/TAP interactive regions of EBNA-1 severely diminished EBNA-1 transactivation of FRTKCAT in transient expression assays. Our data suggest that interaction with P32/TAP may contribute to EBNA-1-mediated transactivation.

Original languageEnglish (US)
Pages (from-to)18-29
Number of pages12
JournalVirology
Volume236
Issue number1
DOIs
StatePublished - Sep 15 1997

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Human Herpesvirus 4
Proteins
Amino Acids
Transcriptional Activation
Cytoplasm
EBV-encoded nuclear antigen 1
Two-Hybrid System Techniques
Epstein-Barr Virus Infections
Viral Genome
Protein C
Glycine
Arginine
Organism Cloning
Binding Sites
Maintenance
Viruses

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Wang, Y., Finan, J. E., Middeldorp, J. M., & Hayward, S. D. (1997). P32/TAP, a cellular protein that interacts with EBNA-1 of Epstein-Barr virus. Virology, 236(1), 18-29. https://doi.org/10.1006/viro.1997.8739

P32/TAP, a cellular protein that interacts with EBNA-1 of Epstein-Barr virus. / Wang, Yilong; Finan, Jon E.; Middeldorp, Jaap M.; Hayward, S. Diane.

In: Virology, Vol. 236, No. 1, 15.09.1997, p. 18-29.

Research output: Contribution to journalArticle

Wang, Y, Finan, JE, Middeldorp, JM & Hayward, SD 1997, 'P32/TAP, a cellular protein that interacts with EBNA-1 of Epstein-Barr virus', Virology, vol. 236, no. 1, pp. 18-29. https://doi.org/10.1006/viro.1997.8739
Wang, Yilong ; Finan, Jon E. ; Middeldorp, Jaap M. ; Hayward, S. Diane. / P32/TAP, a cellular protein that interacts with EBNA-1 of Epstein-Barr virus. In: Virology. 1997 ; Vol. 236, No. 1. pp. 18-29.
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