Site-directed mutagenesis of M1 RNA, the RNA subunit of Escherichia coli ribonuclease P

Lawrence, Nathan P.; Altman, Sidney

doi:10.1016/0022-2836(86)90253-6

Cited by 33 publications

(20 citation statements)

References 27 publications

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“…These data suggest that cells with depleted levels of free C5 protein retained residual RNase P activity, perhaps by some intact M1 RNAs winning in competition with truncated derivatives for C5 protein binding. Alternatively, ribonucleoprotein complexes (composed of truncated M1 RNAs and C5 protein) may have retained some catalytic activities [36,37].…”

Section: Discussionmentioning

confidence: 99%

Novel function of C5 protein as a metabolic stabilizer of M1 RNA

Kim

Lee

2008

FEBS Letters

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Edited by Ulrike KutayKeywords: M1 RNA C5 protein RNase P RNA stability a b s t r a c t Escherichia coli RNase P is a ribonucleoprotein composed of a large RNA subunit (M1 RNA) and a small protein subunit (C5 protein). We examined if C5 protein plays a role in maintaining metabolic stability of M1 RNA. The sequestration of C5 protein available for M1 RNA binding reduced M1 RNA stability in vivo, and its reduced stability was recovered via overexpression of C5 protein. In addition, M1 RNA was rapidly degraded in a temperature-sensitive C5 protein mutant strain at non-permissive temperatures. Collectively, our results demonstrate that the C5 protein metabolically stabilizes M1 RNA in the cell.

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Section: Discussionmentioning

confidence: 99%

Novel function of C5 protein as a metabolic stabilizer of M1 RNA

Kim

Lee

2008

FEBS Letters

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“…This structure, rather than Watson-Crick base pairing with the substrates, is suspected of being primarily responsible for recognition of pre-tRNAs, since several RNA structural disruptions have been shown to reduce catalytic activity. A sequence originally thought to contribute to substrate recognition through complementarity to the invariant T4JCG sequence in tRNAs is dispensable (18,34).RNase P RNA activities have been reported in a number of eucaryotes (1,5,6,8,11,13,22,28,32,33,35,36), but there has been significant disagreement as to the nature of the enzymes. In particular, the RNase P from Xenopis laevis has been reported to be insensitive to micrococcal nuclease and found as part of a large polypeptide complex (6).…”

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confidence: 99%

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confidence: 99%

Partial characterization of an RNA component that copurifies with Saccharomyces cerevisiae RNase P.

Lee¹,

Engelke²

1989

Mol. Cell. Biol.

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Saccharomyces cerevisiae cellular RNase P is composed of both protein and RNA components that are essential for activity. The isolated holoenzyme contains a highly structured RNA of 369 nucleotides that has extensive sequence similarities to the 286-nucleotide RNA associated with Schizosaccharomyces pombe RNase P but bears little resemblance to the analogous RNA sequences in procaryotes or S. cerevisiae mitochondria. Even so, the predicted secondary structure of S. cerevisiae RNA is strikingly similar to the bacterial phylogenetic consensus rather than to previously predicted structures of other eucaryotic RNase P RNAs.Mature 5' termini of tRNAs in both procaryotes and eucaryotes are generated by cleavage with a structurespecific endonuclease, RNase P. Extensive study of RNase P activities from Escherichia coli and Bacillus siubtilis has shown that both enzymes consist of single RNA and protein species (14,16,19,21,25,[29][30][31]38,43,44,51). Although temperature-sensitive mutations in these individual components have shown that both are required for RNase P activity in vivo (24,25,31,38,41,46,47), each RNA component alone can recognize and cleave substrates under some conditions in vitro (2,16,19,20,44). The enzymic RNAs can form extensively base-paired secondary structures as judged by theoretical calculations and cleavage sensitivity structure analyses (3,17,43,44). Extensive phylogenetic sequence comparisons among the bacterial RNAs suggest several short domains of conserved sequences and a conserved secondary structure (26). This structure, rather than Watson-Crick base pairing with the substrates, is suspected of being primarily responsible for recognition of pre-tRNAs, since several RNA structural disruptions have been shown to reduce catalytic activity. A sequence originally thought to contribute to substrate recognition through complementarity to the invariant T4JCG sequence in tRNAs is dispensable (18,34).RNase P RNA activities have been reported in a number of eucaryotes (1,5,6,8,11,13,22,28,32,33,35,36), but there has been significant disagreement as to the nature of the enzymes. In particular, the RNase P from Xenopis laevis has been reported to be insensitive to micrococcal nuclease and found as part of a large polypeptide complex (6). For all other nuclear and mitochondrial enzymes tested, RNA components have been shown to be essential on the basis of either nuclease sensitivity of the enzyme or genetic evidence. Other than one report on HeLa cell RNase P (15), the ability to denature and reconstitute the eucaryotic components has not been reported and there have been no instances of RNA components having catalytic activity in the absence of protein (39). RNA subunits essential for the activity of the enzyme (28, 37) in SchiZosaccharomnvces pombe and Saccharomvces cerev'isiae mitochondria have been identified and their sequences have been determined (33,37). Surprisingly, neither the nucleotide sequences nor the predicted secondary structures of these RNAs bear any striking resemblance to each o...

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“…On the basis of these results we suggest that the mature part of the precursor tRNA may be bound to the central structural domain of M1 RNA which is similar to the central part of the tRNA binding center of 16 S rRNA. Indeed, deletions and mutations in the postulated tRNA binding domain affect the activity of the M1 RNA [13,14].…”

Section: 'mentioning

confidence: 99%

“…the helix 1 (base paired 5'-and 3'-ends of M1 RNA), the helix 2 and the domain built by helices 6-8. From deletion and point mutation experiments Altman and his group concluded recently [13,14] that in M1 RNA there is a separate catalytic center that includes nucleotides 165-255. In the model predicted here for the M1 RNA (figs 1,2) the mentioned sequence region is folded in a discrete domain which contains the helices 6-8 and their adjacent loop regions.…”

Section: 'mentioning

confidence: 99%

Similarities between a predicted secondary structure for the M1 RNA ribozyme and the tRNA binding center of 16 S rRNA from E. coli

Boehm

1987

FEBS Letters

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We propose a new model for the secondary structure of the M ! RNA component of E. coli RNase P which is based on significant sequence homologies with parts of the E. coli 16 S rRNA. A large domain of the new model resembles closely the secondary structure of the tRNA binding center of 16 S rRNA. We suggest that this domain of M 1 RNA when functioning as a ribozyme binds the mature part of the precursor tRNA.

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Site-directed mutagenesis of M1 RNA, the RNA subunit of Escherichia coli ribonuclease P

Cited by 33 publications

References 27 publications

Novel function of C5 protein as a metabolic stabilizer of M1 RNA

Novel function of C5 protein as a metabolic stabilizer of M1 RNA

Partial characterization of an RNA component that copurifies with Saccharomyces cerevisiae RNase P.

Similarities between a predicted secondary structure for the M1 RNA ribozyme and the tRNA binding center of 16 S rRNA from E. coli

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