A homodimer interface without base pairs in an RNA mimic of red fluorescent protein

Nat Chem Biol. 2017 Nov;13(11):1195-1201. doi: 10.1038/nchembio.2475. Epub 2017 Sep 25.

Abstract

Corn, a 28-nucleotide RNA, increases yellow fluorescence of its cognate ligand 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO) by >400-fold. Corn was selected in vitro to overcome limitations of other fluorogenic RNAs, particularly rapid photobleaching. We now report the Corn-DFHO co-crystal structure, discovering that the functional species is a quasisymmetric homodimer. Unusually, the dimer interface, in which six unpaired adenosines break overall two-fold symmetry, lacks any intermolecular base pairs. The homodimer encapsulates one DFHO at its interprotomer interface, sandwiching it with a G-quadruplex from each protomer. Corn and the green-fluorescent Spinach RNA are structurally unrelated. Their convergent use of G-quadruplexes underscores the usefulness of this motif for RNA-induced small-molecule fluorescence. The asymmetric dimer interface of Corn could provide a basis for the development of mutants that only fluoresce as heterodimers. Such variants would be analogous to Split GFP, and may be useful for analyzing RNA co-expression or association, or for designing self-assembling RNA nanostructures.

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Aptamers, Nucleotide / genetics
  • Base Pairing
  • Base Sequence
  • Dimerization*
  • Fluorescent Dyes / metabolism*
  • G-Quadruplexes*
  • Humans
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / metabolism
  • Nucleic Acid Conformation*
  • Optical Imaging / methods*
  • Red Fluorescent Protein

Substances

  • Aptamers, Nucleotide
  • Fluorescent Dyes
  • Luminescent Proteins