Affinity electrophoresis for monitoring terminal phosphorylation and the presence of queuosine in RNA. Application of polyacrylamide containing a covalently bound boronic acid

Nucleic Acids Res. 1985 Oct 11;13(19):6881-98. doi: 10.1093/nar/13.19.6881.

Abstract

An affinity electrophoretic method has been developed to study the state of terminal phosphorylation of RNAs and the presence of the hypermodified base Q in tRNA. It is based on the copolymerization of acryloylaminophenylboronic acid into standard polyacrylamide gels and the interaction of this derivative with free cis-diol groups present in the RNA. In the case of terminal phosphorylation, free ribose groups are present either as such, or may be introduced by enzymatic reactions specific for a particular phosphorylation pattern (e.g. using T4 RNA ligase or guanylyltransferase). Additionally, tRNA species containing the Q base may be resolved from Q-lacking tRNAs by boronate affinity electrophoresis. The introduction of a non-destructive, one-step electrophoretic procedure not only offers an alternative to classical analytical methods, but also provides a means of isolating such populations of RNAs for which other methods are unavailable or are less convenient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamide
  • Acrylamides
  • Boronic Acids*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Guanosine / analogs & derivatives*
  • Nucleoside Q / analysis*
  • Phosphates / analysis*
  • Phosphorylation
  • RNA / analysis*
  • RNA Caps / analysis
  • RNA Processing, Post-Transcriptional

Substances

  • Acrylamides
  • Boronic Acids
  • Phosphates
  • RNA Caps
  • Guanosine
  • Acrylamide
  • Nucleoside Q
  • RNA