Trypanosoma brucei RNA editing complex: band II is structurally critical and maintains band V ligase, which is nonessential

Mol Cell Biol. 2003 Nov;23(21):7909-19. doi: 10.1128/MCB.23.21.7909-7919.2003.

Abstract

Maturation of Trypanosoma brucei mitochondrial mRNA involves massive posttranscriptional insertion and deletion of uridine residues. This RNA editing utilizes an enzymatic complex with seven major proteins, band I through band VII. We here use RNA interference (RNAi) to examine the band II and band V proteins. Band II is found essential for viability; it is needed to maintain the normal structure of the editing complex and to retain the band V ligase protein. Previously, band III was found essential for certain activities, including maintenance of the editing complex and retention of the band IV ligase protein. Thus, band II and band V form a protein pair with features analogous to the band III/band IV ligase pair. Since band V is specific for U insertion and since band IV is needed for U deletion, their parallel organization suggests that the editing complex has a pseudosymmetry. However, unlike the essential band IV ligase, RNAi to band V has only a morphological but no growth rate effect, suggesting that it is stimulatory but nonessential. Indeed, in vitro analysis of band V RNAi cell extract demonstrates that band IV can seal U insertion when band V is lacking. Thus, band IV ligase is the first activity of the basic editing complex shown able to serve in both forms of editing. Our studies also indicate that the U insertional portion may be less central in the editing complex than the corresponding U deletional portion.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Genes, Protozoan
  • Macromolecular Substances
  • Mitochondria / genetics*
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism
  • Molecular Sequence Data
  • Polynucleotide Ligases / genetics
  • Polynucleotide Ligases / metabolism*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism
  • RNA Editing*
  • RNA Interference
  • RNA, Protozoan / metabolism*
  • Sequence Alignment
  • Trypanosoma brucei brucei / cytology
  • Trypanosoma brucei brucei / genetics
  • Trypanosoma brucei brucei / metabolism*

Substances

  • Macromolecular Substances
  • Mitochondrial Proteins
  • Protozoan Proteins
  • RNA, Protozoan
  • Polynucleotide Ligases
  • band V ligase