Ribonuclease P of Tetrahymena thermophila.
Ribonuclease P (RNase P) is responsible for the generation of mature 5' termini of tRNA. The RNA component of this complex encodes the enzymatic activity in bacteria and is itself catalytically active under appropriate conditions in vitro. The role of the subunits in eucaryotes has not yet been established. We have partially purified RNase P activity from the ciliate protozoan Tetrahymena thermophila to learn more about the biochemical characteristics of RNase P from a lower eucaryote. The Tetrahymena RNase P displays a pH optimum and temperature optimum characteristic of RNase P enzymes isolated from other organisms. The Km of the T. thermophila enzyme for pre-tRNAGln is 1.6 x 10(-7)M, which is comparable to the values reported for other examples of RNase P. The Tetrahymena RNase P is a ribonucleoprotein complex, as supported by its sensitivity to micrococcal nuclease and proteinase K. The buoyant density of the enzyme in Cs2SO4 is 1.42 g/ml, which suggests that the RNA component of the Tetrahymena enzyme comprises a significantly greater percentage of the holoenzyme than that determined for RNase P of other Eucarya or Archaea. The holoenzyme has a requirement for divalent cations displaying characteristics that are unique for RNase P but closely resemble preferences reported for the Tetrahymena group I intron RNA. Puromycin inhibits pre-tRNA processing by the Tetrahymena complex, and implications of the similarities between recognition of tRNA by ribosomal components and RNase P are discussed.
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Department of Microbiology and College of Medicine, University of Illinois, Urbana, Illinois 61801, USA.
Celander
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DW
D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D017931:DNA Primers; D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011691:Puromycin / Q000494:pharmacology; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism; D016337:RNA, Catalytic / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012351:RNA, Transfer, Gln / Q000737:chemistry / Q000378:metabolism; D043262:Ribonuclease P; D013379:Substrate Specificity; D016808:Tetrahymena thermophila / Q000201:enzymology*
D002413:Cations, Divalent; D017931:DNA Primers; D012322:RNA Precursors; D016337:RNA, Catalytic; D012351:RNA, Transfer, Gln; D011691:Puromycin; D004722:Endoribonucleases; D043262:Ribonuclease P