Key result
Recombinant REX1 and REL1 successfully reconstitute precleaved U-deletion RNA editing in vitro.
The study demonstrates that REX1 and RNA editing ligase 1 are sufficient to reconstitute precleaved U-deletion RNA editing in vitro.
Defines minimal in vitro editing machinery; leaves open in vivo validation and any therapeutic relevance.
Uridine insertion/deletion RNA editing in trypanosomatid mitochondria is a posttranscriptional RNA modification phenomenon required for translation of mitochondrial mRNAs. This process involves guide RNA-mediated cleavage at a specific site, insertion or deletion of Us from the 3' end of the 5' mRNA fragment, and ligation of the two mRNA fragments. The Leishmania major RNA ligase-containing complex protein 2 expressed in insect cells has a 3'-5' exoribonuclease activity and was therefore renamed RNA editing exonuclease 1 (REX1). Recombinant REX1 specifically trims 3' overhanging Us and stops at a duplex region. Evidence is presented that REX1 is responsible for deletion of the 3' overhanging Us from the bridged mRNA 5' cleavage fragment and that RNA editing ligase 1 is responsible for the ligation of the two mRNA cleavage fragments in U-deletion editing. The evidence involves both in vivo down-regulation of REX1 expression in Trypanosoma brucei by RNA interference and the reconstitution of precleaved U-deletion in vitro editing with only two recombinant enzymes: recombinant REX1 and recombinant RNA editing ligase 1.
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Kang et al. (2005) studied this question. Recombinant REX1 and REL1 was evaluated on Reconstitution of precleaved U-deletion in vitro editing. Recombinant REX1 and RNA editing ligase 1 (REL1) are sufficient to reconstitute precleaved U-deletion RNA editing in vitro.
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