An imperfect correlation between DNA replication activity of Epstein-Barr virus nuclear antigen 1 (EBNA1) and binding to the nuclear import receptor, Rch1/importin alpha

Virology
A L KimPaul M Lieberman

Abstract

Epstein-Barr virus (EBV) replicates as a stable multicopy episome in latently infected mammalian cells. Latent cycle DNA replication requires only two viral elements, the cis-acting origin of plasmid replication (oriP) and the trans-acting origin binding protein (EBNA1). EBNA1 binds multiple recognition sites in oriP, but has not other enzymatic activities associated with replication functions. To identify human cellular proteins that mediate EBNA1 function, we designed a one-hybrid assay in yeast to select for proteins that bind to EBNA1 when bound to criP in vivo. A human cDNA encoding the Rch1/hSRP1 alpha/ importin alpha protein was isolated and shown to bind to full-length EBNA1, but not to an amino terminal deletion mutant of EBNA1 when bound to oriP in yeast. The interaction of EBNA1 with Rch1 was confirmed biochemically by coimmunoprecipitation from nuclear extracts and by direct binding of recombinant proteins in vitro. Internal deletion mutations in EBNA1 which compromised DNA replication activity were similarly reduced for binding to Rch1. Mutations with no effect on DNA replication activity were similarly unaffected for Rch1 binding. Rch1/importin alpha has been shown to bind to the nuclear localization sequence (NLS...Continue Reading

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Citations

Apr 15, 2011·Journal of Virology·Jayaraju Dheekollu, Paul M Lieberman
May 9, 2000·Journal of Virology·D F Ceccarelli, L Frappier
Nov 5, 2002·Journal of Virology·Kong-Nan Zhao, Ian H Frazer
Aug 30, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sibille HummeAloys Schepers
Jul 26, 2003·Folia Microbiologica·J Rajcáni, M Kúdelová
May 25, 2010·Biochemical and Biophysical Research Communications·George KleinElena Kashuba
Jan 24, 2015·Experimental & Molecular Medicine·Myung-Soo Kang, Elliott Kieff
Jan 1, 2012·Scientifica·Lori Frappier
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May 15, 2009·The Journal of General Virology·Sarah J DuellmanRichard R Burgess
Apr 5, 2000·The Journal of Biological Chemistry·C R KovacB K Birshtein
Mar 30, 2001·The Journal of Biological Chemistry·D J Goodwin, A Whitehouse

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