Stability and cooperativity of individual tertiary contacts in RNA revealed through chemical denaturation

Corie Y. Ralston, Qin He, Michael D. Brenowitz, Mark R. Chance

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

For proteins, understanding tertiary interactions involved in local versus global unfolding has become increasingly important for understanding the nature of the native state ensemble, the mechanisms of unfolding, and the stability of both the native and intermediate states in folding. In this work we have addressed related questions with respect to RNA structure by combining chemical denaturation and hydroxyl radical foot-printing methods. We have determined unfolding isotherms for each of 26 discrete sites of protection located throughout the Tetrahymena thermophila group I ribozyme. The cooperativity of folding, m-value, and the free energy, ΔG°(N-U), associated with formation of each tertiary contact was determined by analysis of the isotherms. The ΔG°(N-U) values measured in this study vary from 1.7 ± 0.2 to 7.6 ± 1.2 kcal mol+1. Thus, the stability of these discrete tertiary contacts vary by almost 104. In addition, an intradomain contact and three interdomain contacts show high cooperativity (m-values of 1.1 ± 0.2 to 1.7 ± 0.3 kcal mol+1 M+1) indicating that these contacts exhibit global cooperatively in their folding behavior. This new approach to examining RNA stability provides an exciting comparison to our understanding of protein structure and folding mechanisms.

Original languageEnglish (US)
Pages (from-to)371-374
Number of pages4
JournalNature Structural Biology
Volume7
Issue number5
DOIs
StatePublished - May 2000

Fingerprint

Tetrahymena thermophila
Printing
Denaturation
Protein Folding
RNA Stability
Hydroxyl Radical
Foot
RNA
Isotherms
Proteins
Free energy
GIR1 ribozyme

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology
  • Genetics

Cite this

Stability and cooperativity of individual tertiary contacts in RNA revealed through chemical denaturation. / Ralston, Corie Y.; He, Qin; Brenowitz, Michael D.; Chance, Mark R.

In: Nature Structural Biology, Vol. 7, No. 5, 05.2000, p. 371-374.

Research output: Contribution to journalArticle

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