Comparison of the Structures and Mechanisms of the Pistol and Hammerhead Ribozymes

Timothy J. Wilson, Yijin Liu, Nan-Sheng Li, Qing Dai, Joseph A. Piccirilli, David M. J. Lilley (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    40 Citations (Scopus)
    159 Downloads (Pure)

    Abstract

    Comparison of the secondary and three-dimensional structures of the hammerhead and pistol ribozymes reveals many close similarities, so in this work we have asked if they are mechanistically identical. We have determined a new crystal structure of the pistol ribozyme and have shown that G40 acts as general base in the cleavage reaction. The conformation in the active site ensures an in-line attack of the O2′ nucleophile, and the conformation at the scissile phosphate and the position of the general base are closely similar to those in the hammerhead ribozyme. However, the two ribozymes differ in the nature of the general acid. 2′-Amino substitution experiments indicate that the general acid of the hammerhead ribozyme is the O2′ of G8, while that of the pistol ribozyme is a hydrated metal ion. The two ribozymes are related but mechanistically distinct.

    Original languageEnglish
    Pages (from-to)7865-7875
    Number of pages11
    JournalJournal of the American Chemical Society
    Volume141
    Issue number19
    Early online date24 Apr 2019
    DOIs
    Publication statusPublished - 15 May 2019

    Keywords

    • RNA catalysis
    • general acid-base catalysis
    • metal ions
    • atomic mutagenesis
    • X-ray crystallography

    ASJC Scopus subject areas

    • General Chemistry
    • Biochemistry
    • Catalysis
    • Colloid and Surface Chemistry

    Fingerprint

    Dive into the research topics of 'Comparison of the Structures and Mechanisms of the Pistol and Hammerhead Ribozymes'. Together they form a unique fingerprint.

    Cite this