Characteristics of Ribonuclease P from Various Organisms

Lawrence, Nathan P.; Wesolowski, Donna; Gold, H.; Bartkiewicz, Marcjanna; Guerrier-Takada, Cecilia; McClain, William H.; Altman, Sidney

doi:10.1101/sqb.1987.052.01.028

Cited by 39 publications

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“…The biochemical properties of the two holoenzymes differ substantially, suggesting rather different organizations to the enzymes. The activity of the H. volcanii enzyme, for instance, is reportedly sensitive to treatment with MN, as are bacterial RNase P holoenzymes (124). Similarly, the H. volcanii holoenzyme exhibits a buoyant density of 1.61 g/ml in Cs 2 SO 4 (E. coli RNase P is 1.55 g/ml in Cs 2 SO 4 ), suggestive of a ribonucleoprotein complex consisting primarily of RNA (124).…”

Section: Archaeal Holoenzymesmentioning

confidence: 97%

“…The activity of the H. volcanii enzyme, for instance, is reportedly sensitive to treatment with MN, as are bacterial RNase P holoenzymes (124). Similarly, the H. volcanii holoenzyme exhibits a buoyant density of 1.61 g/ml in Cs 2 SO 4 (E. coli RNase P is 1.55 g/ml in Cs 2 SO 4 ), suggestive of a ribonucleoprotein complex consisting primarily of RNA (124). The RNase P of S. acidocaldarius, in contrast, is both insensitive to MN and has a buoyant density of 1.27 g/ml in Cs 2 SO 4 (125), indicating a considerable protein content.…”

Section: Archaeal Holoenzymesmentioning

confidence: 99%

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RIBONUCLEASE P: Unity and Diversity in a tRNA Processing Ribozyme

Frank

Pace²

1998

Annu. Rev. Biochem.

423

345

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Ribonuclease P (RNase P) is the endoribonuclease that generates the mature 5 -ends of tRNA by removal of the 5 -leader elements of precursor-tRNAs. This enzyme has been characterized from representatives of all three domains of life (Archaea, Bacteria, and Eucarya) (1) as well as from mitochondria and chloroplasts. The cellular and mitochondrial RNase Ps are ribonucleoproteins, whereas the most extensively studied chloroplast RNase P (from spinach) is composed solely of protein. Remarkably, the RNA subunit of bacterial RNase P is catalytically active in vitro in the absence of the protein subunit (2). Although RNA-only activity has not been demonstrated for the archaeal, eucaryal, or mitochondrial RNAs, comparative sequence analysis has established that these RNAs are homologous (of common ancestry) to bacterial RNA. RNase P holoenzymes vary greatly in organizational complexity across the phylogenetic domains, primarily because of differences in the RNase P protein subunits: Mitochondrial, archaeal, and eucaryal holoenzymes contain larger, and perhaps more numerous, protein subunits than do the bacterial holoenzymes. However, that the nonbacterial RNase P RNAs retain significant structural similarity to their catalytically active bacterial counterparts indicates that the RNA remains the catalytic center of the enzyme. CONTENTS

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Section: Archaeal Holoenzymesmentioning

confidence: 97%

Section: Archaeal Holoenzymesmentioning

confidence: 99%

RIBONUCLEASE P: Unity and Diversity in a tRNA Processing Ribozyme

Frank

Pace²

1998

Annu. Rev. Biochem.

423

345

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“…Among the criteria used to elucidate the subunit composition of RNase P from various organisms, the density in isopyknic gradients and the behaviour of the enzyme after treatment with micrococcal nuclease have most often been used to assess the presence of an RNA subunit. Wheat germ RNase P has a density in CsCl which is only slightly above the bulk of protein, but still within the range of RNA enzymes: RNase P from archaebacteria (Lawrence et al, 1987;Darr et al, 1990), yeast mitochondria (Morales et al, 1989), vertebrate cell nuclei (Bartkiewicz et al, 1989;Dona et al, 1991 ;Jayanthi and Van Tuyle, 1992) and the photosynthetic organelles of Cyanophora parudoxa (Cordier, A. and Schon, A., unpublished results) have densities ranging between 1.27-1.61 g/ml in Cs,SO,, yet they all contain RNAs of comparable size (ibid. ;and Nieuwlandt et al, 1991;Altman et al, 1993b;LaGrandeur et al, 1993;Baum et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Partial Purification and Characterization of Nuclear Ribonuclease P from Wheat

Arends

Schön

1997

European Journal of Biochemistry

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Ribonuclease P (RNase P) from wheat nuclei has been purified over 1000-fold, using wheat germ extract as starting material and a combination of poly(ethylenglyco1) precipitation and column chromatography. The enzyme was shown to be of nuclear origin by its characteristic ionic requirements; for optimum activity it requires 0.5-1.5 mM Mg", which can be partly replaced by MnZ+. With about 100 kDa, wheat nuclear RNase P has the lowest molecular mass reported so far for a eukaryotic RNase P. The enzyme has an isoelectric point of 5.0 and a buoyant density of 1.34 g/ml in CsCl, suggesting the presence of a nucleic acid component; it is, however, insensitive against treatment with micrococcal nuclease. Wheat germ RNase P requires an intact tertiary structure of the pre-tRNA substrate; its cleavage efficiency is also influenced by the presence of an intron, and by the nature of the 3' terminus of the substrate. The apparent K,,, and V,,, for an intronless plant pre-tRNATYr are 10.3 nM and 1.12 fmol/min, respectively.Keywords: enzyme purification; nuclear ribonuclease P; pre-tRNA processing ; substrate specificity ; Triticum aestivum.RNase P is the ubiquitous endonuclease required for the maturation of the 5' end of tRNAs. In all cases where the composition of this enzyme has been investigated in detail, it was found to consist of both protein and RNA subunits (Kmpp et al., 1986;Bartkiewicz et al., 1989;Lee and Engelke, 1989;Morales et al., 1989;Doria et al., 1991;Nieuwlandt et al., 1991 ; LaGrandeur et al., 1993;Marchfelder and Brennicke, 1994;Baum et al., 1996; Lee et al., 1996a,b; for the bacterial enzymes, see reviews by Altman et al., 1993a;Pace and Brown, 1995). RNase P RNAs from bacteria show enzymatic activity in vitro without the protein subunit; however, RNase P from archaebacteria or eukaryotes is only active as a holoenzyme. In contrast to our current knowledge on RNase P from bacteria and most eukaryotic cellular compartments, no detailed information is available on cleavage mechanism, substrate requirements, or subunit composition of RNase P from plant nuclei. Previous studies performed with a complete processing system from wheat germ did not allow unambigous determination of the order of cleavage events involved in end maturation of tRNAs (e.g. Stange and Beier, 1987). Similarly, although investigation of a variety of pre-tRNAs indicated that mutations disrupting the tRNA structure severely impair overall processing in this plant in vitro system (Stange et al., 1991 ;Fuchs et al., 1992;Teichmann et al., 1994), it is not clear at present which of the enzymes involved is most sensitive to these changes. To approach these questions, and to gain insight into the molecular composition and substrate requirements of nuclear RNase P from higher (EC 3.1.31.1).plants, we have purified and characterized this enzyme from wheat germ. Although an RNA component could not yet be identified in wheat germ RNase P, several properties of the enzyme indicate that it might be a ribonucleoprotein. The availability of a high...

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“…The H. volcanii RNase P, on the other hand, resembles the bacterial enzyme in density in Cs 2 SO 4 (1.61 g͞cm 3 ; ref. 10) and nuclease sensitivity. The H. volcanii enzyme contains a 435-nt RNA (8).…”

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confidence: 99%

RNase P RNAs from some Archaea are catalytically active

Pannucci

Haas

Hall

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

202

152

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The RNA subunits of RNase Ps of Archaea and eukaryotes have been thought to depend fundamentally on protein for activity, unlike those of Bacteria that are capable of efficient catalysis in the absence of protein. Although the eukaryotic RNase P RNAs are quite different than those of Bacteria in both sequence and structure, the archaeal RNAs generally contain the sequences and structures of the bacterial, phylogenetically conserved catalytic core. A spectrum of archaeal RNase P RNAs were therefore tested for activity in a wide range of conditions. Many remain inactive in ionically extreme conditions, but catalytic activity could be detected from those of the methanobacteria, thermococci, and halobacteria. Chimeric holoenzymes, reconstituted from the Methanobacterium RNase P RNA and the Bacillus subtilis RNase P protein subunits, were functional at low ionic strength. The properties of the archaeal RNase P RNAs (high ionic-strength requirement, low affinity for substrate, and catalytic reconstitution by bacterial RNase P protein) are similar to synthetic RNase P RNAs that contain all of the catalytic core of the bacterial RNA but lack phylogenetically variable, stabilizing elements.RNase P is an endoribonuclease best known for its role in tRNA biosynthesis, in which it is the enzyme responsible for the removal of 5Ј leader sequences from transfer RNA precursors (for reviews see refs. 1 and 2). In Bacteria, RNase P consists of two subunits: a large (Ϸ140-kDa, 400-nt) RNA and a small (Ϸ14-kDa, 120-amino acid) protein. Both RNA and protein are required in vivo and for optimal activity in vitro in reactions at low ionic strength (3, 4). The RNA is the catalytic subunit of the bacterial enzyme; at elevated ionic strength in vitro, it is by itself capable of processing pre-tRNAs catalytically; i.e., it is a ribozyme (5). The protein component of the bacterial enzyme alters substrate recognition by directly contacting the leader region of the pre-tRNA (6). The RNase P enzymes of Archaea (formerly archaebacteria) and Eukarya (the nuclear͞cytoplasmic portion of the eukaryotic cell) also contain RNA subunits, but these RNAs have not been shown to be catalytically active. Although it seems likely that the catalytic function of the archaeal and eukaryal enzymes resides in the RNA, the expression of this activity apparently requires the presence of the protein subunits.RNase P enzymes have been characterized from only two archaeal species: the thermoacidophilic crenarchaeote Sulfolobus acidocaldarius (7) and the extremely halophilic euryarchaeote Haloferax volcanii (8). The S. acidocaldarius RNase P is resistant to micrococcal nuclease treatment and contains a 315-nt RNA that persists after nuclease treatment (9). The enzyme is large (Ϸ400 kDa apparent molecular mass) and has a low density in Cs 2 SO 4 (1.27 g͞cm 3 ), similar to the densities of the eukaryotic nuclear enzyme and implying a high protein:RNA content. The H. volcanii RNase P, on the other hand, resembles the bacterial enzyme in density in Cs 2 SO 4 (1.61...

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Characteristics of Ribonuclease P from Various Organisms

Cited by 39 publications

References 0 publications

RIBONUCLEASE P: Unity and Diversity in a tRNA Processing Ribozyme

RIBONUCLEASE P: Unity and Diversity in a tRNA Processing Ribozyme

Partial Purification and Characterization of Nuclear Ribonuclease P from Wheat

RNase P RNAs from some Archaea are catalytically active

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