Rational optimization of the DSL ligase ribozyme with GNRA/receptor interacting modules

Arch Biochem Biophys. 2009 Oct 15;490(2):163-70. doi: 10.1016/j.abb.2009.08.020. Epub 2009 Sep 1.

Abstract

The DSL ribozyme is a class of artificial ligase ribozymes with a highly modular architecture, which catalyzes template-directed RNA ligation on a helical substrate module that can be either covalently connected (cis-DSL) or physically separated (trans-DSL) from the catalytic module. Substrate recognition by the catalytic module is promoted by one or two sets of GNRA/receptor interactions acting as clamps in the cis or trans configurations, respectively. In this study, we have rationally designed and analyzed the catalytic and self-assembly properties of several trans-DSL ribozymes with different sets of natural and artificial GNRA-receptor clamps. Two variants newly designed in this study showed significantly enhanced catalytic properties with respect of the original trans-DSL construct. While this work allows dissection of the turnover and catalytic properties of the trans-DSL ribozyme, it also emphasizes the remarkable modularity of RNA tertiary structure for nano-construction of complex functions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Biomedical Engineering
  • Catalytic Domain
  • Drug Design
  • Kinetics
  • Nucleic Acid Conformation
  • RNA / genetics
  • RNA / metabolism
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • Stereoisomerism
  • Substrate Specificity

Substances

  • RNA, Catalytic
  • RNA