title: Characterization of ribonuclease P from the archaebacterium Sulfolobus solfataricus.
abstract: Ribonuclease P is the endonuclease that removes the leader fragments from the 5'-ends of precursor tRNAs. The enzyme isolated from eubacteria contains a catalytic RNA subunit. RNAs also copurify with eukaryotic RNase P, although catalysis by those RNAs has not been demonstrated. This paper reports the isolation and characterization of ribonuclease P from the thermoacidophilic archaebacterium Sulfolobus solfataricus. Archaebacteria are a primary evolutionary lineage, distinct from both eukaryotes and eubacteria. Ribonuclease P of S. solfataricus has reaction component requirements and a Km for substrate tRNA (2.5 X 10(-7) M) that are roughly similar to those reported for eubacterial and eukaryotic ribonuclease P. The temperature optimum for the reaction is 77 degrees C, reflecting the thermophilic character of the organism. The enzyme activity is not affected by treatment with micrococcal nuclease, suggesting that there is no RNA subunit or that it is protected from nuclease action. The density of the enzyme in cesium sulfate equilibrium density gradients is 1.27 g/ml, which is similar to that of protein. However, several RNAs between 200 and 400 nucleotides in size copurify with the enzyme activity on the density gradients, and one of them remains after micrococcal nuclease treatment. These properties of the S. solfataricus enzyme are compared with those of ribonuclease P from eukaryotes and eubacteria.
authors.affiliation: Department of Biology, Indiana University, Bloomington 47405.
authors.forename: S C
authors.initials: SC
authors.lastname: Darr
authors.forename: B
authors.initials: B
authors.lastname: Pace
authors.forename: N R
authors.initials: NR
authors.lastname: Pace
mesh_terms: D001105:Archaea / Q000201:enzymology*; D001419:Bacteria / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D004722:Endoribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D012322:RNA Precursors / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D043262:Ribonuclease P
chemical_list: D012322:RNA Precursors; D012343:RNA, Transfer; D004722:Endoribonucleases; D043262:Ribonuclease P