title: tRNA 3' processing in plants: nuclear and mitochondrial activities differ.
abstract: The nuclear tRNA 3' processing activity from wheat has been characterized and partially purified. Several characteristics of the wheat nuclear 3' processing enzyme now allow this activity to be distinguished from its mitochondrial counterpart. The nuclear enzyme is an endonuclease, which we termed nuclear RNase Z. The enzyme cleaves at the discriminator base and seems to consist only of protein subunits, since essential RNA subunits could not be detected. RNase Z leaves 5' terminal phosphoryl and 3' terminal hydroxyl groups at the processing products. It is a stable enzyme being active over broad temperature and pH ranges, with the highest activity at 35 degrees C and pH 8.4. The apparent molecular mass according to gel filtration chromatography is 122 kDa. The nuclear RNase Z does process 5' extended pretRNAs but with a much lower efficiency than 5' matured pretRNAs. Nuclear intron-containing precursor tRNAs as well as mitochondrial precursor tRNAs are efficiently cleaved by the nuclear RNase Z. Mitochondrial pretRNA(His) is processed by the nuclear RNase Z, generating a mature tRNA(His) containing an 8 base pair acceptor stem. The edited mitochondrial pretRNA(Phe) is cleaved easily, while the unedited version having a mismatch in the acceptor stem is not cleaved. Thus, an intact acceptor stem seems to be required for processing. Experiments with precursors containing mutated tRNAs showed that a completely intact anticodon arm is not necessary for processing by RNase Z. Comparison of the plant nuclear tRNA 3' processing enzyme with the plant mitochondrial one suggests that both activities are different enzymes.
authors.affiliation: Allgemeine Botanik, Universität Ulm, Albert-Einstein-Allee 11, D-89069 Ulm, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Mayer
authors.forename: S
authors.initials: S
authors.lastname: Schiffer
authors.forename: A
authors.initials: A
authors.lastname: Marchfelder
mesh_terms: D002467:Cell Nucleus / Q000235:genetics*; D004722:Endoribonucleases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007438:Introns; D008928:Mitochondria / Q000235:genetics*; D008957:Models, Genetic; D016296:Mutagenesis; D017393:RNA Editing; D012322:RNA Precursors / Q000378:metabolism*; D018749:RNA, Plant / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity; D014908:Triticum / Q000201:enzymology / Q000235:genetics
chemical_list: D012322:RNA Precursors; D018749:RNA, Plant; D012343:RNA, Transfer; D004722:Endoribonucleases; C110250:RNase Z