A Modular RNA Domain That Confers Differential Ligand Specificity

Biochemistry. 2020 Apr 7;59(13):1361-1366. doi: 10.1021/acs.biochem.0c00117. Epub 2020 Mar 27.

Abstract

The modularity of protein domains is well-known, but the existence of independent domains that confer function in RNA is less established. Recently, a family of RNA aptamers termed ykkC was discovered; they bind at least four ligands of very different chemical composition, including guanidine, phosphoribosyl pyrophosphate (PRPP), and guanosine tetraphosphate (ppGpp) (graphical abstract). Structures of these aptamers revealed an architecture characterized by two coaxial helical stacks. The first helix appears to be a generic scaffold, while the second helix forms the most contacts to the ligands. To determine if these two regions within the aptamer are modular units for ligand recognition, we swapped the ligand-binding coaxial stacks of a guanidine aptamer and a PRPP aptamer. This operation, in combination with a single mutation in the scaffold domain, achieved full switching of ligand specificity. This finding suggests that the ligand-binding helix largely dictates the ligand specificity of ykkC RNAs and that the scaffold coaxial stack is generally compatible with various ykkC ligand-binding modules. This work presents an example of RNA domain modularity comparable to that of a ligand-binding protein, showcasing the versatility of RNA as an entity capable of molecular evolution through adaptation of existing motifs.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / metabolism*
  • Guanosine Tetraphosphate / chemistry
  • Guanosine Tetraphosphate / metabolism
  • Ligands
  • Models, Molecular
  • Nucleic Acid Conformation
  • Phosphoribosyl Pyrophosphate / chemistry
  • Phosphoribosyl Pyrophosphate / metabolism

Substances

  • Aptamers, Nucleotide
  • Ligands
  • Guanosine Tetraphosphate
  • Phosphoribosyl Pyrophosphate