Nucleic Acids Research, 1993, Vol. 21, No. 19 4483-4490
© 1993
RNA |
Comparative analysis of the regulation of the interferoninducible protein kinase PKR by Epstein - Barr virus RNAs EBER-1 and EBER-2 and adenovirus VA, RNA
1Institut for Biologie III, Universitat Freiburg D-7800 Freiburg, Germany 2Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk Bristol BS8 1TD, UK Division of Biochemistry, Department of Cellular and Molecular Sciences, St George's Hospital Medical School Cranmer Terrace, London SW17 ORE, UK
*To whom correspondence should be addressed
Received June 24, 1993. Revised August 9, 1993. Accepted August 9, 1993.
The interferon-inducible protein kinase PKR interacts with a number of small viral RNA species, including adenovirus VA, RNA and the Epstein-Barr virusencoded RNA EBER-1. These RNAs bind to PKR and protect protein synthesis from inhibition by doublestranded RNA in the reticulocyte lysate system. Using a peptide phosphorylation assay we show here that EBER-1, like VA,, directly inhibits the activation of purified PKR. A second Epstein-Barr virus RNA, EBER-2, also regulates PKR. EBER-1, EBER-2 and VA, RNA exhibit mutually competitive binding to the native or recombinant enzyme, as assessed by U.V. crosslinking experiments and filter binding assays. The affinities of all three RNAs for PKR in vitro are similar (Kd = ca. 0.3 nM). Since this protein kinase has been proposed to exert a tumour suppressor function in vivo, the ability of EBER-1 to inhibit its activation suggests a role for this small RNA in cell transformation by Epstein - Barr virus.
+Persent addresses: Department of Virology, University of Freiburg, Hermann-Herder-Strasse 11, D-7800 Freiburg, Germany.
Department of Cellular and Molecular Physiology, Milton S.Hershey Medical Center, PO Box 850, Hershey, PA 17033, USA.
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