Processing of transcripts of a dimeric tRNA gene in yeast uses the nuclease responsible for maturation of the 3′ termini upon 5 S and 37 S precursor rRNAs

Piper, Peter W.; Stråby, Kerstin B.

doi:10.1016/0014-5793(89)80745-8

Cited by 18 publications

(15 citation statements)

References 17 publications

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“…Strain PP1002, however, harbors an rna82 lesion that causes aberrant 3' end processing in 5S and tRNA RNA polymerase III transcripts (46)(47)(48). In this strain, a faint signal corre- sponding to an additional 28 to 30 nucleotides past the 369-nucleotide signal was seen in some but not all experiments.…”

Section: Resultsmentioning

confidence: 90%

Characterization of RPR1, an essential gene encoding the RNA component of Saccharomyces cerevisiae nuclear RNase P.

Lee¹,

Rohlman²,

Molony³

et al. 1991

Mol. Cell. Biol.

115

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RNA components have been identified in preparations of RNase P from a number of eucaryotic sources, but final proof that these RNAs are true RNase P subunits has been elusive because the eucaryotic RNAs, unlike the procaryotic RNase P ribozymes, have not been shown to have catalytic activity in the absence of protein.We previously identified such an RNA component in Saccharomyces cerevisiae nuclear RNase P preparations and have now characterized the corresponding, chromosomal gene, called RPRI (RNase P ribonucleoprotein 1). Gene disruption experiments showed RPR1 to be single copy and essential. Characterization of the gene region located RPRI 600 bp downstream of the URA3 coding region on chromosome V. We have sequenced 400 bp upstream and 550 bp downstream of the region encoding the major 369-nucleotide RPRI RNA. The presence of less abundant, potential precursor RNAs with an extra 84 nucleotides of 5' leader and up to 30 nucleotides of 3' trailing sequences suggests that the primary RPRI transcript is subjected to multiple processing steps to obtain the 369-nucleotide form. Complementation of RPRI-disrupted haploids with one variant of RPRI gave a slow-growth and temperature-sensitive phenotype. This strain accumulates tRNA precursors that lack the 5' end maturation performed by RNase P, providing direct evidence that RPRI RNA is an essential component of this enzyme.RNase P from both procaryotic and eucaryotic sources is an endonuclease that cleaves pre-tRNA substrates to yield mature 5' termini. In procaryotes, both the RNA and protein components of the ribonucleoprotein enzymes are required for in vivo activity (15,27,29,33,43,49,57,58), but the RNA component alone is capable of efficiently catalyzing the correct reaction under some conditions in vitro (3,18,21,22,52). Extensive phylogenetic sequence comparisons of these RNAs (30), combined with folding energy calculations and cleavage sensitivity studies (6, 19, 51, 52), suggest a conserved, highly ordered secondary structure. Most if not all of the key contacts with the substrates depend on this structure, with the protein contributing to efficiency through secondary effects such as charge shielding between RNA chain phosphate backbones. The mechanism by which pretRNA substrates are recognized is not clearly understood, however, since there are no obvious regions of required Watson-Crick base pairing between the enzyme and substrate RNAs (20,36).Studies of eucaryotic RNase Ps have suggested that they also contain essential RNA subunits, although the role of the RNA components has not been firmly established. RNAs that copurify with both nuclear and organelle RNase Ps have been characterized (1,7,9,11,12,14,24,32,34,35,37,38,40,42) and shown to have sequence and structural similarities to each other and to a lesser degree to the procaryotic RNAs (5, 37). The identification of the eucaryotic RNAs as essential RNase P subunits has not been rigorously confirmed, however, since there are no reports of the eucaryotic RNAs having enzyme activity in the absence ...

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Section: Resultsmentioning

confidence: 90%

Characterization of RPR1, an essential gene encoding the RNA component of Saccharomyces cerevisiae nuclear RNase P.

Lee¹,

Rohlman²,

Molony³

et al. 1991

Mol. Cell. Biol.

115

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“…39 End processing of pre-tRNAs generally follows 59 end processing, and is catalyzed by both exonucleases (Rex1 in yeast) (Piper and Straby 1989;Copela et al 2008;Ozanick et al 2009) and the RNase Z endonuclease (Trz1 in yeast) (for review, see Vogel et al 2005). The balance between endonucleolytic and exonucleolytic 39 end processing depends, at least in part, on the tRNA-binding La protein in both lower and higher eukaryotes Copela et al 2008;Zhao et al 2009).…”

Section: Org Downloaded Frommentioning

confidence: 99%

tRNA biology charges to the front

Phizicky¹,

Hopper²

2010

Genes Dev.

686

687

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tRNA biology has come of age, revealing an unprecedented level of understanding and many unexpected discoveries along the way. This review highlights new findings on the diverse pathways of tRNA maturation, and on the formation and function of a number of modifications. Topics of special focus include the regulation of tRNA biosynthesis, quality control tRNA turnover mechanisms, widespread tRNA cleavage pathways activated in response to stress and other growth conditions, emerging evidence of signaling pathways involving tRNA and cleavage fragments, and the sophisticated intracellular tRNA trafficking that occurs during and after biosynthesis.

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“…Previously described nuclear 3'-processing activities from several organisms except the yeast S. cerevisiae all cleaved their substrates between bases 73 and 74 of the mature tRNA domain. In contrast, the S. cerevisiae 3'-processing enzyme is an exonuclease (12,33). We were curious to learn whether plant nuclear tRNA 3'-processing activity was exo-or endonucleolytic and, if it was endonucleolytic, whether it would cleave at the same site as the other nuclear activities or at the site chosen by the chloroplast enzyme.…”

Section: )mentioning

confidence: 99%

“…In most well-characterized eukaryotic tRNA processing systems, the 3' cleavage enzyme is an endonuclease; this has been conclusively demonstrated in extracts of human (1,46), Xenopus laevis (6,16,23), and Drosophila (14) cells, mitochondria from rat liver (28) and yeast (7) cells, and spinach chloroplasts (44). Only in yeast nuclei are some tRNA trailers known to be removed by the action of a 3'-5' exonuclease (12,33). The tRNA 3' endonucleases from human, X. laevis, and Drosophila cells and from yeast and rat liver mitochondria are specific for 5'-mature forms of those pre-tRNAs which were tested (6,7,14,28,46).…”

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confidence: 98%

Cleavage specificity of chloroplast and nuclear tRNA 3'-processing nucleases.

Oommen

Gegenheimer

1992

Mol. Cell. Biol.

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tRNAs in eukaryotic nuclei and organelles are synthesized as precursors lacking the 3'-terminal CCA sequence and possessing 5' (leader) and 3' (trailer) extensions. Nucleolytic cleavage of the 3' trailer and addition of CCA are therefore required for formation of functional tRNA 3' termini. Many chloroplast tRNA genes encode a C at position 74 which is not removed during processing but which can be incorporated as the first base of the CCAOH terminus. Sequences downstream of nucleotide 74, however, are always removed.Synthetic yeast pre-tRNA"te substrates containing the complete CCA7476 sequence were processed with crude or partially purified chloroplast enzyme fractions. The 3'-extended substrates (tRNA-CCA-trailer) were cleaved exclusively between nucleotides 74 and 75 to give tRNA-COH, whereas a 3'-mature transcript (tRNA-CCAoH) was not cleaved at all. A 5'-, 3'-extended chloroplast tRNA-CAG-trailer was also processed entirely to tRNA-COH. Furthermore, a 5'-mature, 3'-extended yeast pre-tRNAPhe derivative, tRNA-ACAtrailer, in which C74 was replaced by A, was cleaved precisely after A74. In contrast, we found that a partially purified enzyme fraction (a nuclear/cytoplasmic activity) from wheat embryo cleaved the 3'-extended yeast tRNAPhe precursors between nucleotides 73 and 74 to give tRNAOH. This specificity is consistent with that of all previously characterized nuclear enzyme preparations. We conclude that (i) chloroplast tRNA 3'-processing endonuclease cleaves after nucleotide 74 regardless of the nature of the surrounding sequences; (ii) this specificity differs from that of the plant nuclear/cytoplasmic processing nuclease, which cleaves after base 73; and (iii) since 3'-mature tRNA is not a substrate for either activity, these 3' nucleases must require substrates possessing a 3'-terminal extension that extends past nucleotide 76. This substrate specificity may prevent mature tRNA from counterproductive cleavage by the 3' processing system.

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Processing of transcripts of a dimeric tRNA gene in yeast uses the nuclease responsible for maturation of the 3′ termini upon 5 S and 37 S precursor rRNAs

Cited by 18 publications

References 17 publications

Characterization of RPR1, an essential gene encoding the RNA component of Saccharomyces cerevisiae nuclear RNase P.

Characterization of RPR1, an essential gene encoding the RNA component of Saccharomyces cerevisiae nuclear RNase P.

tRNA biology charges to the front

Cleavage specificity of chloroplast and nuclear tRNA 3'-processing nucleases.

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