tRNA 3' end maturation in archaea has eukaryotic features: the RNase Z from Haloferax volcanii.
Here, we report the first characterization and partial purification of an archaeal tRNA 3' processing activity, the RNase Z from Haloferax volcanii. The activity identified here is an endonuclease, which cleaves tRNA precursors 3' to the discriminator. Thus tRNA 3' processing in archaea resembles the eukaryotic 3' processing pathway. The archaeal RNase Z has a KCl optimum at 5mM, which is in contrast to the intracellular KCl concentration being as high as 4M KCl. The archaeal RNase Z does process 5' extended and intron-containing pretRNAs but with a much lower efficiency than 5' matured, intronless pretRNAs. At least in vitro there is thus no defined order for 5' and 3' processing and splicing. A heterologous precursor tRNA is cleaved efficiently by the archaeal RNase Z. Experiments with precursors containing mutated tRNAs revealed that removal of the anticodon arm reduces cleavage efficiency only slightly, while removal of D and T arm reduces processing effciency drastically, even down to complete inhibition. Comparison with its nuclear and mitochondrial homologs revealed that the substrate specificity of the archaeal RNase Z is narrower than that of the nuclear RNase Z but broader than that of the mitochondrial RNase Z.
Schierling
Karina
K
Molekulare Botanik, Universität Ulm, Germany.
Rösch
Sylvia
S
Rupprecht
Renate
R
Schiffer
Steffen
S
Marchfelder
Anita
A
D000926:Anticodon / Q000235:genetics; D001483:Base Sequence; D002467:Cell Nucleus / Q000201:enzymology; D004722:Endoribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000201:enzymology*; D019143:Evolution, Molecular; D019622:Haloferax volcanii / Q000201:enzymology* / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D007438:Introns / Q000235:genetics; D008928:Mitochondria / Q000201:enzymology; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology; D026721:RNA 3' End Processing; D019642:RNA, Archaeal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012365:RNA, Transfer, Tyr / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D000926:Anticodon; D019642:RNA, Archaeal; D012365:RNA, Transfer, Tyr; D011189:Potassium Chloride; D012343:RNA, Transfer; D004722:Endoribonucleases; C110250:RNase Z; C045782:3'-pre-tRNase