Transfer RNA intron processing in the halophilic archaebacteria.
An in vitro assay system has been developed for the Halobacterium volcanii tRNA intron endonuclease using in vitro generated precursor RNAs. A partially purified enzyme preparation is capable of precise and accurate excision of the intron from the halobacterial tRNA(Trp) precursor. The cleavage reaction produces products having 5' hydroxyl and 2',3' cyclic phosphate termini. Processing of precursor molecules containing deletions within the exon regions indicates that the halobacterial endonuclease does not require intact mature tRNA structure in the substrate; this is in contrast to the eukaryotic endonuclease enzyme that has an absolute requirement for these structures. The large halobacterial tRNA(Trp) intron does not appear to be a primary site for recognition by the endonuclease, however, its removal affects cleavage efficiency. Through a comparison of the structural and sequence features of the halobacterial substrates and the precursors of other archaebacterial intron-containing precursors, a common element is proposed for the recognition of substrates by intron endonuclease.
Thompson
L D
LD
Department of Microbiology, Ohio State University, Columbus 43210.
Brandon
L D
LD
Nieuwlandt
D T
DT
Daniels
C J
CJ
D001483:Base Sequence; D004722:Endoribonucleases / Q000378:metabolism; D005798:Genes, Bacterial; D006217:Halobacterium / Q000235:genetics / Q000378:metabolism*; D007438:Introns; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
D012322:RNA Precursors; D012329:RNA, Bacterial; D012343:RNA, Transfer; D004722:Endoribonucleases; C057798:tRNA(Trp) intron endonuclease