Comparison of the hammerhead cleavage reactions stimulated by monovalent and divalent cations

Jessica L. O'Rear, Shenglong Wang, Andrew L. Feig, Leonid Beigelman, Olke C. Uhlenbeck, Daniel Herschlag*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

157 Scopus citations

Abstract

Although the hammerhead reaction proceeds most efficiently in divalent cations, cleavage in 4 M LiCl is only ∼10-fold slower than under standard conditions of 10 mM MgCl2 (Murray et al., Chem Biol, 1998, 5:587-595; Curtis & Bartel, RNA, 2001, this issue, pp. 546-552). To determine if the catalytic mechanism with high concentrations of monovalent cations is similar to that with divalent cations, we compared the activities of a series of modified hammerhead ribozymes in the two ionic conditions. Nearly all of the modifications have similar deleterious effects under both reaction conditions, suggesting that the hammerhead adopts the same general catalytic structure with both monovalent and divalent cations. However, modification of three ligands previously implicated in the binding of a functional divalent metal ion have substantially smaller effects on the cleavage rate in Li+ than in Mg2+. This result suggests that an interaction analogous to the interaction made by this divalent metal ion is absent in the monovalent reaction. Although the contribution of this divalent metal ion to the overall reaction rate is relatively modest, its presence is needed to achieve the full catalytic rate. The role of this ion appears to be in facilitating formation of the active structure, and any direct chemical role of metal ions in hammerhead catalysis is small.

Original languageEnglish (US)
Pages (from-to)537-545
Number of pages9
JournalRNA
Volume7
Issue number4
DOIs
StatePublished - 2001

Keywords

  • Biological catalysis
  • Enzyme
  • Mechanistic analysis
  • Metal ions
  • RNA cleavage
  • Thio effects

ASJC Scopus subject areas

  • Molecular Biology

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