TY - JOUR
T1 - 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
AU - Sharp, Tyson V.
AU - Schwemmle, Martin
AU - Jeffrey, Ian
AU - Laing, Kenneth
AU - Mellor, Harry
AU - Proud, Christopher G.
AU - Hilse, Kurt
AU - Clemens, Michael J.
N1 - Funding Information:
We are most grateful to Drs. Glen Barber and Michael Katze (University of Washington, Seattle) and Dr Dirk Gewert (Wellcome Foundation, Beckenham) for assistance in production of recombinant PKR in the baculovirus-infected insect cell system. This work was funded by grants from the Cancer Research Campaign (SP1951/0501), the Leukaemia Research Fund (91/59) and the Wellcome Trust (031488) (to M.J.C.), the Medical Research Council (to C.G.P.) and the Deutsche Forschungsgemeinschaft (to K.H.). T.S. is supported by a studentship from the Science and Engineering Research Council and M.S. received a graduate fellowship from the State of Baden Wurttemberg. Exchange of personnel between the UK and Germany was facilitated by a grant from the Anglo-German Academic Research Collaboration Programme.
PY - 1993/9/25
Y1 - 1993/9/25
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0027487288&partnerID=8YFLogxK
U2 - 10.1093/nar/21.19.4483
DO - 10.1093/nar/21.19.4483
M3 - Article
C2 - 7901835
AN - SCOPUS:0027487288
SN - 0305-1048
VL - 21
SP - 4483
EP - 4490
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 19
ER -