The genomic HDV ribozyme utilizes a previously unnoticed U-turn motif to accomplish fast site-specific catalysis

Nucleic Acids Res. 2007;35(6):1933-46. doi: 10.1093/nar/gkl1104. Epub 2007 Mar 2.

Abstract

The genome of the human hepatitis delta virus (HDV) harbors a self-cleaving catalytic RNA motif, the genomic HDV ribozyme, whose crystal structure shows the dangling nucleotides 5' of the cleavage site projecting away from the catalytic core. This 5'-sequence contains a clinically conserved U-1 that we find to be essential for fast cleavage, as the order of activity follows U-1 > C-1 > A-1 > G-1, with a >25-fold activity loss from U-1 to G-1. Terbium(III) footprinting detects conformations for the P1.1 stem, the cleavage site wobble pair and the A-minor motif of the catalytic trefoil turn that depend on the identity of the N-1 base. The most tightly folded catalytic core, resembling that of the reaction product, is found in the U-1 wild-type precursor. Molecular dynamics simulations demonstrate that a U-1 forms the most robust kink around the scissile phosphate, exposing it to the catalytic C75 in a previously unnoticed U-turn motif found also, for example, in the hammerhead ribozyme and tRNAs. Strikingly, we find that the common structural U-turn motif serves distinct functions in the HDV and hammerhead ribozymes.

Publication types

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

MeSH terms

  • Base Sequence
  • Binding Sites
  • Catalysis
  • Computer Simulation
  • Genome, Viral
  • Hepatitis Delta Virus / enzymology*
  • Hepatitis Delta Virus / genetics
  • Hydrogen Bonding
  • Magnesium / chemistry
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Purines / chemistry
  • Pyrimidines / chemistry
  • RNA, Catalytic / chemistry*
  • RNA, Viral / chemistry*
  • Terbium / chemistry
  • Uracil / chemistry

Substances

  • Purines
  • Pyrimidines
  • RNA, Catalytic
  • RNA, Viral
  • Terbium
  • Uracil
  • Magnesium