Identification of a 90-kDa polypeptide which associates with adenovirus VA RNAI and is phosphorylated by the double-stranded RNA-dependent protein kinase

Rice, A. P., Kostura, M., Mathews, M. B. (December 1989) Identification of a 90-kDa polypeptide which associates with adenovirus VA RNAI and is phosphorylated by the double-stranded RNA-dependent protein kinase. J Biol Chem, 264 (34). pp. 20632-7. ISSN 0021-9258

Abstract

Interferon treatment of mammalian cells induces a double-stranded (ds) RNA-dependent protein kinase known as DAI. When activated, DAI phosphorylates the alpha-subunit of eukaryotic initiation factor eIF-2, impairing its ability to be recycled and leading to the inhibition of protein synthesis. We have identified a novel DAI substrate in the ribosomal salt wash of rabbit reticulocyte lysates. This substrate is a 90-kDa polypeptide which has been purified to apparent homogeneity. It can be cross-linked by ultraviolet irradiation to adenovirus VA RNAI, a small RNA polymerase III transcript RNA which acts as an inhibitor of DAI. As assayed by a nitrocellulose filter binding assay, the 90-kDa polypeptide is also able to associate with authentic double-stranded RNA, but not single-stranded RNA, made in vitro. Thus, this newly identified substrate of DAI appears to have affinity for dsRNA structures and may be involved in dsRNA-regulated processes in the reticulocyte. Polyclonal and monoclonal antibodies directed against the 90-kDa polypeptide co-precipitate DAI, suggesting that these two proteins may exist as a complex.

Item Type: Paper
Uncontrolled Keywords: Adenoviruses, Human/*metabolism Animals Antibodies Antigen-Antibody Complex/analysis Electrophoresis, Polyacrylamide Gel Hela Cells/metabolism Humans Molecular Weight Phosphorylation Protein Kinases/*metabolism RNA, Viral/biosynthesis/isolation & purification/*metabolism Rabbits Research Support, U.S. Gov't, P.H.S. Reticulocytes/metabolism Viral Proteins/isolation & purification/*metabolism eIF-2 Kinase
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
organism description > virus > adenovirus
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > polypeptides
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date: 5 December 1989
Date Deposited: 27 Jun 2017 16:03
Last Modified: 27 Jun 2017 16:03
Related URLs:
URI: https://repository.cshl.edu/id/eprint/34898

Actions (login required)

Administrator's edit/view item Administrator's edit/view item