Selection of high affinity RNA ligands to the bacteriophage R17 coat protein

J Mol Biol. 1992 Dec 5;228(3):862-9. doi: 10.1016/0022-2836(92)90870-p.

Abstract

RNA ligands with high affinity for the bacteriophage R17 coat protein were isolated from a pool of random RNA molecules using SELEX. Of the 38 ligands isolated, 36 were found to contain a hairpin very similar to the naturally occurring coat protein binding site in the R17 genome. The common features of these 36 sequences provide a consensus binding site and predict components of a hairpin that promote favorable interaction with the coat protein. These include a tetraloop of primary sequence AUCA and a variable-length stem with a bulged adenosine residue at a specific stem position. The predicted consensus agrees well with the highest-affinity RNA binding site of the R17 coat protein, identified through classical but laborious techniques. These results demonstrate the value of SELEX as a tool for isolating high affinity RNA ligands to a specific target protein, and the further value of those ligands to point the researcher toward natural sequences for that target protein.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Allosteric Regulation
  • Bacteriophages / chemistry*
  • Bacteriophages / ultrastructure
  • Base Sequence
  • Binding Sites
  • Capsid / metabolism*
  • Consensus Sequence
  • Electrophoresis / methods
  • Isomerism
  • Ligands
  • Macromolecular Substances
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Oligoribonucleotides / metabolism*
  • Sequence Homology, Nucleic Acid

Substances

  • Ligands
  • Macromolecular Substances
  • Oligoribonucleotides