Chimeric gRNA-mRNA molecules with oligo(U) tails covalently linked at sites of RNA editing suggest that U addition occurs by transesterification

• Published in: Cell Vol. 65; no. 4; pp. 543 - 550
• Main Authors: Blum, Beat, Sturm, Nancy R., Simpson, Agda M., Simpson, Larry
• Format: Journal Article
• Language: English
• Published: Cambridge, MA Elsevier Inc 17.05.1991; Cell Press
• Subjects: Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Cells, Cultured; Chimera; Cloning, Molecular; Electron Transport Complex IV - genetics; Fundamental and applied biological sciences. Psychology; Macromolecular Substances; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; NADH Dehydrogenase - genetics; Nucleic Acid Conformation; Nucleic Acid Hybridization; Oligonucleotide Probes; Oligoribonucleotides - metabolism; Polymerase Chain Reaction; RNA - genetics; RNA, Guide; RNA, Messenger - genetics; Transcription. Transcription factor. Splicing. Rna processing; Uracil Nucleotides - metabolism; Leishmania tarentolae; Northern blotting; Protozoa; RNA editing; Molecular processing; Messenger RNA; Rhizoflagellata; Models; Transesterification; Nucleic acid; Mechanism; Uracil nucleotide
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/0092-8674(91)90087-F

Chimeric RNA molecules were detected by polymerase chain reaction ampllfication 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 resuit 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.