Perturbation of the hierarchical folding of a large RNA by the destabilization of its scaffold's tertiary structure

Inna Shcherbakova, Michael D. Brenowitz

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The P4-P6 domain serves as a scaffold against which the periphery and catalytic core organize and fold during Mg2+-mediated folding of the Tetrahymena thermophila ribozyme. The most prominent structural motif of the P4-P6 domain is the tetraloop-tetraloop receptor interaction which "clamps" the distal parts of its hairpin-like structure. Destabilization of the tertiary structure of the P4-P6 domain by perturbation of the tetraloop-tetraloop receptor interaction alters the Mg2+- mediated folding pathway. The folding hierarchy of P5c≈P4- P6>periphery>catalytic core that is a striking attribute of the folding of the wild-type RNA is abolished. The initial steps in folding of the mutant RNA are ≥50-fold faster than those of the wild-type ribozyme with the earliest observed tertiary contacts forming around regions known to specifically bind Mg2+. The interaction between the mutant tetraloop and the tetraloop receptor appears coincidently with slowly forming catalytic core tertiary contacts. Thus, the stability conferred upon the P4-P6 domain by the tetraloop-tetraloop receptor interaction dictates the preferred folding pathway by stabilizing an early intermediate. A sub-denaturing concentration of urea diminishes the early barrier to folding the wild-type ribozyme along with complex effects on the subsequent steps of folding the wild-type and mutant RNA.

Original languageEnglish (US)
Pages (from-to)483-496
Number of pages14
JournalJournal of Molecular Biology
Volume354
Issue number2
DOIs
StatePublished - Nov 25 2005

Fingerprint

Catalytic RNA
RNA Folding
Catalytic Domain
RNA
Tetrahymena thermophila
Urea

Keywords

  • Cations
  • Cynchrotron
  • Folding
  • RNA
  • Tetrahymena

ASJC Scopus subject areas

  • Virology

Cite this

Perturbation of the hierarchical folding of a large RNA by the destabilization of its scaffold's tertiary structure. / Shcherbakova, Inna; Brenowitz, Michael D.

In: Journal of Molecular Biology, Vol. 354, No. 2, 25.11.2005, p. 483-496.

Research output: Contribution to journalArticle

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