The cellular protein PRA1 modulates the anti-apoptotic activity of Epstein-Barr virus BHRF1, a homologue of Bcl-2, through direct interaction.
The Epstein-Barr virus-encoded early protein, BHRF1, is a structural and functional homologue of the anti-apoptotic protein, Bcl-2. There is accumulating evidence that BHRF1 protects a variety of cell types from apoptosis induced by various external stimuli. To identify specific proteins from normal epithelial cells that interact with BHRF1 and that might promote or inhibit its anti-apoptotic activity, we screened a yeast two-hybrid cDNA library derived from human normal foreskin keratinocytes and identified a cellular gene encoding human prenylated rab acceptor 1 (hPRA1). The interaction of hPRA1 with BHRF1 was confirmed using glutathione S-transferase pull-down assays, confocal laser scanning microscopy, and co-immunoprecipitation. Two regions of PRA1, amino acids 30-53 and the carboxyl-terminal 21 residues, are important for BHRF1 interactions and two regions of BHRF1, amino acids 1-18 and 89-142, including the Bcl-2 homology domains BH4 and BH1, respectively, are crucial for PRA1 interactions. PRA1 expression interferes with the anti-apoptotic activity of BHRF1, although not of Bcl-2. These results indicate that the PRA1 interacts selectively with BHRF1 to reduce its anti-apoptotic activity and might play a role in the impeding completion of virus maturation.
Li
L Y
LY
Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
Shih
H M
HM
Liu
M Y
MY
Chen
J Y
JY
D017209:Apoptosis / Q000502:physiology*; D002352:Carrier Proteins; D002460:Cell Line; D005656:Fungal Proteins / Q000378:metabolism / Q000502:physiology*; D019204:GTP-Binding Proteins; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins; D011233:Precipitin Tests; D011485:Protein Binding; D011956:Receptors, Cell Surface; D033921:Vesicular Transport Proteins; D014764:Viral Proteins / Q000378:metabolism / Q000502:physiology*
C083278:BHRF1 protein, Human herpesvirus 4; D002352:Carrier Proteins; D005656:Fungal Proteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D033921:Vesicular Transport Proteins; D014764:Viral Proteins; D019204:GTP-Binding Proteins; C108941:RABAC1 protein, human