A re-investigation of the thio effect at the hammerhead cleavage site

Edmund C. Scott, Olke C. Uhlenbeck*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

95 Scopus citations

Abstract

The effect of introducing a phosphorothioate at the hammerhead cleavage site was investigated using a kinetically well-characterized hammerhead. In buffers containing Mg ion, the Rp-phosphorothioate isomer cleaved 2000- to 80,000-fold slower than the Sp isomer or the unmodified RNA substrate. Addition of low concentrations of several thiophilic metal ions, especially Cd2+, to these reactions is sufficient to fully restore the cleavage rate of the Rp substrate without affecting cleavage rate of the all-oxygen or Sp substrate. Thus, a model proposing coordination of a divalent metal ion to the pro-R oxygen at the hammerhead cleavage site appears justified.

Original languageEnglish (US)
Pages (from-to)479-484
Number of pages6
JournalNucleic acids research
Volume27
Issue number2
DOIs
StatePublished - Jan 15 1999

Funding

This work was supported by grant GM 36944.

ASJC Scopus subject areas

  • Genetics

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