title: KREX2 is not essential for either procyclic or bloodstream form Trypanosoma brucei.
abstract: BACKGROUND
  Most mitochondrial mRNAs in Trypanosoma brucei require RNA editing for maturation and translation. The edited RNAs primarily encode proteins of the oxidative phosphorylation system. These parasites undergo extensive changes in energy metabolism between the insect and bloodstream stages which are mirrored by alterations in RNA editing. Two U-specific exonucleases, KREX1 and KREX2, are both present in protein complexes (editosomes) that catalyze RNA editing but the relative roles of each protein are not known.
  
  
  RESULTS
  The requirement for KREX2 for RNA editing in vivo was assessed in both procyclic (insect) and bloodstream form parasites by methods that use homologous recombination for gene elimination. These studies resulted in null mutant cells in which both alleles were eliminated. The viability of these cells demonstrates that KREX2 is not essential in either life cycle stage, despite certain defects in RNA editing in vivo. Furthermore, editosomes isolated from KREX2 null cells require KREX1 for in vitro U-specific exonuclease activity.
  
  
  CONCLUSIONS
  KREX2 is a U-specific exonuclease that is dispensable for RNA editing in vivo in T. brucei BFs and PFs. This result suggests that the U deletion activity, which is required for RNA editing, is primarily mediated in vivo by KREX1 which is normally found associated with only one type of editosome. The retention of the KREX2 gene implies a non-essential role or a role that is essential in other life cycle stages or conditions.
authors.affiliation: Seattle Biomedical Research Institute, Seattle, Washington, United States of America.
authors.forename: Jason
authors.initials: J
authors.lastname: Carnes
authors.forename: Nancy
authors.initials: N
authors.lastname: Lewis Ernst
authors.forename: Carey
authors.initials: C
authors.lastname: Wickham
authors.forename: Brian
authors.initials: B
authors.lastname: Panicucci
authors.forename: Kenneth
authors.initials: K
authors.lastname: Stuart
mesh_terms: D000818:Animals; D005092:Exonucleases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D055785:Gene Knockdown Techniques; D017125:Genes, Protozoan; D051379:Mice; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017393:RNA Editing / Q000235:genetics / Q000502:physiology; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D005092:Exonucleases