ELAV proteins stabilize deadenylated intermediates in a novel in vitro mRNA deadenylation/degradation system.

Published

Journal Article

We have developed an in vitro mRNA stability system using HeLa cell cytoplasmic S100 extracts and exogenous polyadenylated RNA substrates that reproduces regulated aspects of mRNA decay. The addition of cold poly(A) competitor RNA activated both a sequence-specific deadenylase activity in the extracts as well as a potent, ATP-dependent ribonucleolytic activity. The rates of both deadenylation and degradation were up-regulated by the presence of a variety of AU-rich elements in the body of substrate RNAs. Competition analyses demonstrated that trans-acting factors were required for RNA destabilization by AU-rich elements. The approximately 30-kD ELAV protein HuR specifically bound to RNAs containing an AU-rich element derived from the TNF-alpha mRNA in the in vitro system. Interaction of HuR with AU-rich elements, however, was not associated with RNA destabilization. Interestingly, recombinant ELAV proteins specifically stabilized deadenylated intermediates generated from the turnover of AU-rich element-containing substrate RNAs. These data suggest that mammalian ELAV proteins play a role in regulating mRNA stability by influencing the access of degradative enzymes to RNA substrates.

Full Text

Duke Authors

Cited Authors

  • Ford, LP; Watson, J; Keene, JD; Wilusz, J

Published Date

  • January 15, 1999

Published In

Volume / Issue

  • 13 / 2

Start / End Page

  • 188 - 201

PubMed ID

  • 9925643

Pubmed Central ID

  • 9925643

International Standard Serial Number (ISSN)

  • 0890-9369

Digital Object Identifier (DOI)

  • 10.1101/gad.13.2.188

Language

  • eng

Conference Location

  • United States