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PMID: 1673026 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Binding of Epstein-Barr virus small RNA EBER-1 to the double-stranded RNA-activated protein kinase DAI.

Nucleic acids research ·Vol. 19 ·No. 2 ·1991-01-25 ·Pages 243-8

Clarke PA, Schwemmle M, Schickinger J, Hilse K, Clemens MJ

Abstract

Epstein-Barr virus encodes two small RNAs, EBER-1 and -2, that are abundantly expressed in latently infected cells. Recent evidence suggests a role for EBER-1 in regulation of translation since this RNA is able to prevent the inhibition of protein synthesis by double-stranded RNA in rabbit reticulocyte lysates. We show here that EBER-1 that has been synthesized in vitro forms a complex with the dsRNA-activated inhibitor of protein synthesis DAI, a protein kinase that specifically phosphorylates polypeptide chain initiation factor eIF-2. Gel retardation assays and UV crosslinking experiments indicate that complex formation is specific for EBER-1 and requires the presence of some secondary structure in the molecule. RNA competition studies show that EBER-1-DAI complex formation is not inhibited in the presence of other small RNA species, heparin or the synthetic double-stranded RNA, poly(I).poly(C). SDS gel analysis reveals the existence of two forms of the crosslinked complex, of 64-68kDa and 46-53kDa, both of which are recognized by anti-DAI antibodies in immunoprecipitation experiments. These data suggest that EBER-1 regulates protein synthesis through its ability to interact with DAI.

MeSH Terms
Blotting, Western Cross-Linking Reagents Electrophoresis, Polyacrylamide Gel Herpesvirus 4, Human/genetics,metabolism Phosphorylation Plasmids Precipitin Tests Protein Biosynthesis Protein Kinases/metabolism RNA Polymerase III/genetics RNA, Double-Stranded/genetics,metabolism RNA, Viral/genetics,metabolism Ultraviolet Rays eIF-2 Kinase
Chemicals
Cross-Linking Reagents RNA, Double-Stranded RNA, Viral Protein Kinases eIF-2 Kinase RNA Polymerase III
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Clarke P A
Department of Cellular and Molecular Sciences, St George's Hospital Medical School, London, UK.
Schwemmle M
Schickinger J
Hilse K
Clemens M J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/19.2.243
Published
1991-01-25
Pages
243-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC333586
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/19.2.243 PMC Full Text View journal details
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