Chimeric gRNA-mRNA molecules with oligo(U) tails covalently linked at sites of RNA editing suggest that U addition occurs by transesterification.
Chimeric RNA molecules were detected by polymerase chain reaction amplification of kinetoplast RNA using a 3' primer specific to mRNA and a 5' primer specific to guide RNA (gRNA), and directly by Northern analysis. Covalent linkage of the 3' oligo(U) tail of the gRNA to the mRNA occurs at editing sites. Chimeric molecules were isolated for NADH dehydrogenase subunit 7 and cytochrome oxidase subunits II and III. We propose that these molecules are intermediates in the editing process and that successive transesterifications result in the transfer of uridine residues from the gRNA 3' oligo(U) tail to an editing site, with the number of uridine residues determined by base pairing with adenine and guanine "guide" nucleotides in the gRNA.
Blum
B
B
Department of Biology, University of California, Los Angeles 90024.
Sturm
N R
NR
Simpson
A M
AM
Simpson
L
L
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D002678:Chimera; D003001:Cloning, Molecular; D003576:Electron Transport Complex IV / Q000235:genetics*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D009843:Oligoribonucleotides / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D014500:Uracil Nucleotides / Q000378:metabolism*; D058727:RNA, Small Untranslated
D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D009843:Oligoribonucleotides; D012333:RNA, Messenger; D014500:Uracil Nucleotides; C027916:oligo(U); D012313:RNA; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV; D058727:RNA, Small Untranslated