RNA splicing in higher plant mitochondria: determination of functional elements in group II intron from a chimeric cox II gene in electroporated wheat mitochondria.
Higher plant mitochondria mainly contain group II introns presenting a secondary structure with six helical domains linked to a central hub. Experimental evidence of functional elements in higher plant mitochondria introns is limited since they are unable to undergo self-splicing and the definition of functional domains is based on data obtained from yeast autocatalytic introns. Here we study the role of putative functional elements required for the splicing reaction. The exon-binding and intron-binding sites (EBS and IBS, respectively), and the domain 6, which is involved in lariat formation, were analysed by site-directed mutagenesis and transient expression in electroporated mitochondria. The data presented here demonstrate the role of EBS1-IBS1 and EBS2-IBS2 interactions and reveal a new secondary-structure interaction. The role of the C to U editing conversion in the IBS1 motif is discussed.
Farré
Jean-Claude
JC
Laboratoire de Réplication et Expression des Gènes Eucaryotes et Rétroviraux, UMR 5097, Centre National de la Recherche Scientifique and Université Victor Segalen-Bordeaux II. 146, rue Leo Saignat. 33076 Bordeaux Cedex, France.
Araya
Alejandro
A
D001483:Base Sequence; D001665:Binding Sites; D002872:Chromosome Deletion; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018274:Electroporation / Q000379:methods; D007438:Introns / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009690:Nucleic Acid Conformation; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics*; D017381:Spliceosomes / Q000235:genetics / Q000378:metabolism; D014908:Triticum / Q000235:genetics* / Q000378:metabolism
D010940:Plant Proteins; D012322:RNA Precursors; C111957:cox2 protein, Glycine soja; D003576:Electron Transport Complex IV