Near native structure in an RNA collapsed state

Biochemistry. 2003 Dec 2;42(47):13869-78. doi: 10.1021/bi035476k.

Abstract

Many large RNAs form conformationally collapsed, but non-native, states prior to folding to the native state or assembling with protein cofactors. Although RNA collapsed states play fundamental roles in RNA folding and ribonucleoprotein assembly processes, their structures have been poorly understood. We obtained 12 high-quality structural constraints for the collapsed state formed by the catalytic core of the bI5 intron RNA using site-specific cross-linking mediated by a short-lived reactant. RNA tertiary structures in the collapsed and native states are indistinguishable, even though only the native state forms a solvent-inaccessible core. Thus, structural neighbors in the collapsed state, including several long-range tertiary interactions, are approximately as close in space as in the native state, but RNA packing is sufficiently loose or dynamic to allow access by solvent. Binding by the obligate CBP2 protein cofactor has almost no effect on structural neighbors reported by cross-linking, even though protein binding chases the RNA from the collapsed state to the native state. Protein binding thus appears to promote only the final few angstroms of RNA folding rather than mediate global conformational rearrangements in the catalytic core. The bI5 RNA collapsed state functions to self-chaperone ribonucleoprotein assembly because this conformationally restrained structure lies very near that of the native state and excludes structures that otherwise misassemble efficiently.

Publication types

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

MeSH terms

  • Azides / chemistry
  • Base Sequence
  • Cross-Linking Reagents / chemistry
  • Cytochromes b / chemistry
  • Cytochromes b / genetics
  • Introns
  • Magnesium / chemistry
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation*
  • Nucleic Acid Heteroduplexes / chemistry
  • Photochemistry
  • Protein Binding / genetics
  • RNA Processing, Post-Transcriptional
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / genetics
  • RNA, Fungal / chemistry
  • Ribonucleoproteins / chemistry
  • Ribonucleoproteins / genetics
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Ultraviolet Rays

Substances

  • Azides
  • CBP2 protein, S cerevisiae
  • Cross-Linking Reagents
  • Nucleic Acid Heteroduplexes
  • RNA, Catalytic
  • RNA, Fungal
  • Ribonucleoproteins
  • Saccharomyces cerevisiae Proteins
  • Cytochromes b
  • Magnesium