High resolution crystal structure of a key editosome enzyme from Trypanosoma brucei: RNA editing ligase 1

J Mol Biol. 2004 Oct 22;343(3):601-13. doi: 10.1016/j.jmb.2004.08.041.

Abstract

Trypanosomatids are causative agents of several devastating tropical diseases such as African sleeping sickness, Chagas' disease and leishmaniasis. There are no effective vaccines available to date for treatment of these protozoan diseases, while current drugs have limited efficacy, significant toxicity and suffer from increasing resistance. Trypanosomatids have several remarkable and unique metabolic and structural features that are of great interest for developing new anti-protozoan therapeutics. One such feature is "RNA editing", an essential process in these pathogenic protozoa. Transcripts for key trypanosomatid mitochondrial proteins undergo extensive post-transcriptional RNA editing by specifically inserting or deleting uridylates from pre-mature mRNA in order to create mature mRNAs that encode functional proteins. The RNA editing process is carried out in a approximately 1.6 MDa multi-protein complex, the editosome. In Trypanosoma brucei, one of the editosome's core enzymes, the RNA editing ligase 1 (TbREL1), has been shown to be essential for survival of both insect and bloodstream forms of the parasite. We report here the crystal structure of the catalytic domain of TbREL1 at 1.2 A resolution, in complex with ATP and magnesium. The magnesium ion interacts with the beta and gamma-phosphate groups and is almost perfectly octahedrally coordinated by six phosphate and water oxygen atoms. ATP makes extensive direct and indirect interactions with the ligase via essentially all its atoms while extending its base into a deep pocket. In addition, the ATP makes numerous interactions with residues that are conserved in the editing ligases only. Further away from the active site, TbREL1 contains a unique loop containing several hydrophobic residues that are highly conserved among trypanosomatid RNA editing ligases which may play a role in protein-protein interactions in the editosome. The distinct characteristics of the adenine-binding pocket, and the absence of any close homolog in the human genome, bode well for the design of selective inhibitors that will block the essential RNA ligase function in a number of major protozoan pathogens.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Carbon-Oxygen Ligases / chemistry*
  • Carbon-Oxygen Ligases / genetics
  • Carbon-Oxygen Ligases / metabolism
  • Catalytic Domain
  • Crystallography, X-Ray
  • Humans
  • Magnesium / metabolism
  • Mitochondrial Proteins / chemistry*
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Multienzyme Complexes
  • Protein Structure, Secondary
  • Protein Structure, Tertiary*
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism
  • RNA Editing*
  • Sequence Alignment
  • Trypanosoma brucei brucei / enzymology*
  • Trypanosoma brucei brucei / genetics

Substances

  • Mitochondrial Proteins
  • Multienzyme Complexes
  • Protozoan Proteins
  • Adenosine Triphosphate
  • Carbon-Oxygen Ligases
  • REL1 RNA ligase, Trypanosoma brucei
  • Magnesium

Associated data

  • PDB/1XDN