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PMID: 8552671 Published · ppublish English Journal Article

In vitro activation of the interferon-induced, double-stranded RNA-dependent protein kinase PKR by RNA from the 3' untranslated regions of human alpha-tropomyosin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 1 ·1996-01-09 ·Pages 508-13

Davis S, Watson JC

Abstract

The cellular kinase known as PKR (protein kinase RNA-activated) is induced by interferon and activated by RNA. PKR is known to have antiviral properties due to its role in translational control. Active PKR phosphorylates eukaryotic initiation factor 2 alpha and leads to inhibition of translation, including viral translation. PKR is also known to function as a tumor suppressor, presumably by limiting the rate of tumor-cell translation and growth. Recent research has shown that RNA from the 3' untranslated region (3'UTR) of human alpha-tropomyosin has tumor-suppressor properties in vivo [Rastinejad, F., Conboy, M. J., Rando, T. A. & Blau, H. M. (1993) Cell 75, 1107-1117]. Here we report that purified RNA from the 3'UTR of human alpha-tropomyosin can inhibit in vitro translation in a manner consistent with activation of PKR. Inhibition of translation by tropomyosin 3'UTR RNA was observed in a rabbit reticulocyte lysate system, which is known to contain endogenous PKR but was not seen in wheat germ lysate, which is not responsive to a known activator of PKR. A control RNA purified in the same manner as the 3'UTR RNA did not inhibit translation in either system. The inhibition of translation observed in reticulocyte lysates was prevented by the addition of adenovirus virus-associated RNA1 (VA RNAI), an inhibitor of PKR activation. Tropomyosin 3'UTR RNA was bound by immunoprecipitated PKR and activated the enzyme in an in vitro kinase assay. These data suggest that activation of PKR could be the mechanism by which tropomyosin 3'UTR RNA exerts its tumor-suppression activity in vivo.

MeSH Terms
Animals Base Sequence Cell-Free System Cloning, Molecular DNA Primers/chemistry Enzyme Activation Humans Molecular Sequence Data Protein Biosynthesis Protein Serine-Threonine Kinases/metabolism RNA, Messenger/metabolism Rabbits Reticulocytes Tropomyosin/genetics eIF-2 Kinase
Chemicals
DNA Primers RNA, Messenger Tropomyosin Protein Serine-Threonine Kinases eIF-2 Kinase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Davis S
RiboGene, Inc., Hayward, CA 94545, USA.
Watson J C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.1.508
Published
1996-01-09
Pages
508-13
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC40267
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.1.508 PMC Full Text View journal details
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