RNA-protein interaction methods to study viral IRES elements

Methods. 2015 Dec:91:3-12. doi: 10.1016/j.ymeth.2015.06.023. Epub 2015 Jul 2.

Abstract

Translation control often takes place through the mRNA untranslated regions, involving direct interactions with RNA-binding proteins (RBPs). Internal ribosome entry site elements (IRESs) are cis-acting RNA regions that promote translation initiation using a cap-independent mechanism. A subset of positive-strand RNA viruses harbor IRESs as a strategy to ensure efficient viral protein synthesis. IRESs are organized in modular structural domains with a division of functions. However, viral IRESs vary in nucleotide sequence, secondary RNA structure, and transacting factor requirements. Therefore, in-depth studies are needed to understand how distinct types of viral IRESs perform their function. In this review we describe methods to isolate and identify RNA-binding proteins important for IRES activity, and to study the impact of RNA structure and RNA-protein interactions on IRES activity.

Keywords: IRES elements; RNA structure; RNA virus; RNA-binding proteins; Translation control.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Chromatography, Affinity / methods
  • Electrophoresis, Capillary
  • Internal Ribosome Entry Sites*
  • Mass Spectrometry
  • Nucleic Acid Conformation
  • RNA Viruses / genetics
  • RNA Viruses / metabolism
  • RNA, Messenger / metabolism
  • RNA, Viral / metabolism*
  • RNA-Binding Proteins / isolation & purification
  • RNA-Binding Proteins / metabolism*

Substances

  • Internal Ribosome Entry Sites
  • RNA, Messenger
  • RNA, Viral
  • RNA-Binding Proteins