Nucleotide sequence of the region between the 18S rRNA sequence and the 28S rRNA sequence of rat ribosomal DNA

Subrahmanyam, C S; Cassidy, B. G.; Busch, Harris; Rothblum, Lawrence I.

doi:10.1093/nar/10.12.3667

Cited by 95 publications

(44 citation statements)

References 24 publications

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“…2). These were noted in a tobacco chloroplast S19 gene (25) and a rat rRNA spacer sequence (24). Whether these sequences participate in mechanisms that regulate the ribosomal gene family and how they might be involved in coordinating the expression of ribosomal protein and rRNA genes is not clear.…”

Section: Discussionmentioning

confidence: 99%

Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs.

Rhoads¹,

Roufa²

1985

Mol. Cell. Biol.

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The Chinese hamster ovary (CHO) cell 40S ribosomal subunit protein S14 provides a unique opportunity to investigate an important mammalian housekeeping gene and its mRNA and protein products. The S14 gene appears to be single copy, and CHO cell S14 mutants have been isolated as emetine-resistant (emtB) clones in tissue culture. Thus, S14 is the only mammalian ribosomal protein whose gene structure and function are amenable to straightforward genetic and biochemical analysis. Recently, we isolated a wild-type Chinese hamster lung cell cDNA clone, pCS14-1, including an almost complete copy of the ribosomal protein S14 message (N. Nakamichi, D. D. Rhoads, and D. J. Roufa, J. Biol. Chem. 258:13236-13242, 1983). Here we describe comparable cDNAs from wild-type and emtB CHO cells. We report both mRNA and polypeptide sequences of the wild-type and mutant ribosomal protein transcripts. As a consequence of the genetic methods used to obtain our emetine-resistant mutants, the emtB S14 cDNAs differ from wild-type cDNA by single-base changes. Physical and chemical features of polypeptides encoded by the cDNAs are consistent with wellcharacterized S14 protein polymorphisms. The three emtB mutations analyzed affect two adjacent arginine codons within the very basic S14 carboxyl region, indicating a significant role for this portion of the protein in the function and architecture of the mammalian 40S ribosomal subunit.Among mammalian ribosomal protein genes, the Chinese hamster ovary (CHO) cell locus specifying 40S subunit protein S14 is particularly suited to direct biochemical and genetic analysis. S14 is the only mammalian ribosomal protein gene for which both mutants and molecular probes are available. We have identified protein S14 as the product of the CHO cell emetine resistance locus emtB (1, 2, 13, 14), and we have described bacterial plasmids containing Chinese hamster lung (CHL) cell S14 cDNAs (18). To characterize this ribosomal protein gene and its emtB alleles, we have used a CHL cDNA (pCS14-1) to isolate wild-type and mutant CHO cell S14 cDNAs.In the present study, we determined the CHO S14 cDNA sequence and precise nucleic acid and protein alterations affected by several emtB mutations. Only a few single-base substitutions distinguish mutant from wild-type cDNAs. Resulting amino acid replacements account for electrophoretic variants of S14 protein associated with emtB mutations (1, 2, 13, 14). Our data indicate that the carboxyl region of the S14 protein is an important determinant of 40S ribosomal subunit structure and function. MATERIALS AND METHODSCell lines, microbial strains, and materials. The wild-type and mutant Chinese hamster cell lines and methods used to maintain them in tissue culture have been described previously (1,2,13,14,18 Unlabeled deoxyribonucleoside, dideoxyribonucleoside triphosphates, and M13 "17mer" primer were obtained from P-L Biochemicals, Inc., Milwaukee, Wis. Restriction endonucleases, oligodeoxyguanylate-tailed, PstI-cleaved pBR322, Escherichia coli DNA polymerase I (Klenow fragme...

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Section: Discussionmentioning

confidence: 99%

Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs.

Rhoads¹,

Roufa²

1985

Mol. Cell. Biol.

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“…In case RNase treatment was performed, cells were incubated with 100 µg/ml RNase A for 1 hour at 37°C and subsequently washed three times with 2× SSC. Digoxigenin-labeled probes to rRNAs, generated by nick translation from plasmids containing parts of the mature regions of 18S and 28S rRNA, respectively Subrahmanyam et al, 1982), were a kind gift of C. Carvalho and M. Carmo-Fonseca (University of Lisbon, Portugal) (Carvalho et al, 2001). Prior to hybridization, cells were washed with 50% deionized formamide, 2× SSC.…”

Section: Molecular Cloning Protein Expression and Purificationmentioning

confidence: 99%

Biogenesis and nuclear export of ribosomal subunits in higher eukaryotes depend on the CRM1 export pathway

Thomas

Kutay

2003

Journal of Cell Science

165

188

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The production of ribosomes constitutes a major biosynthetic task for cells. Eukaryotic small and large ribosomal subunits are assembled in the nucleolus and independently exported to the cytoplasm. Most nuclear export pathways require RanGTP-binding export receptors. We analyzed the role of CRM1, the export receptor for leucine-rich nuclear export signals (NES), in the biogenesis of ribosomal subunits in vertebrate cells. Inhibition of the CRM1 export pathway led to a defect in nuclear export of both 40S and 60S subunits in HeLa cells. Moreover, the export of newly made ribosomal subunits in Xenopus oocytes was efficiently and specifically competed by BSA-NES conjugates. The CRM1 dependence of 60S subunit export suggested a conserved function for NMD3, a factor proposed to be a 60S subunit export adaptor in yeast. Indeed, we observed that nuclear export of human NMD3(hNMD3) is sensitive to leptomycin B (LMB), which inactivates CRM1. It had,however, not yet been demonstrated that Nmd3 can interact with CRM1. Using purified recombinant proteins we have shown here that hNMD3 binds to CRM1 directly, in a RanGTP-dependent manner, by way of a C-terminal NES sequence. Our results suggest that the functions of CRM1 and NMD3 in ribosomal subunit export are conserved from yeast to higher eukaryotes.

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“…The 5' end of the 26S rRNA could not be satisfactorily determined. However, the 5' terminal sequences of 26/28S rRNA have been reported for Xenopus, yeast, rat, and mouse (5,6,(20)(21)(22) and in each case the terminal nucleotide is either 2 or 3 bases 5' to a conserved GACCUC sequence. This sequence, which has been implicated in binding with the 3' end of the 5.8S rRNA (20) is also found in the sea urchin rDNA unit at positions 1100-1105.…”

Section: Theand Mn Teminalnauleotidmentioning

confidence: 99%

“…The complete sequence of the 26S rDNA has been determined for two species of Saccharomyces (20,21) and for Physarum (28); the first 69 nucleotides of rat 28S rDNA (6), the first 118 nucleotides of Xenopus 28S rDNA (5), and the first 585 nucleotides of mouse 28S rDNA (22) have been published. Comparison of these sequences to the first 505 nucleotides of Lytechinus reveals two regions of high homology.…”

Section: Regionmentioning

confidence: 99%

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Nucleotide sequence of the 18S–26S rRNA intergene region of the sea urchin

Hindenach¹,

Stafford²

1984

Nucl Acids Res

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The DNA sequence which spans the internal transcribed spacers of a cloned ribosomal transcription unit from the sea urchin, Lyteehinus variegatus, has been determined. The region extends from the conserved Eco RI site near the 3' end of the 18S rDNA to a Bam HI site in the 26S rDNA and includes 232 nucleotides coding for 18S rRNA, 367 nucleotides of internal transcribed spacer, 159 nucleotides coding for 5.8S rRNA, 338 nucleotides of internal transcribed spacer, and 505 nucleotides coding for 26S rRNA. The rRNA coding regions were identified by direct analysis of 3'-labeled 18S and 5.8S rRNA and 5'-labeled 5. . INTBODUCTLIQNEukaryotic ribosomal RNA is synthesized as a large precursor containing 17/18S, 5.8S, and 26/28S rRNAs plus spacer sequences. The RNA is posttranscriptionally modified and cleaved at specific sites to produce a set of intermediate precursors and finally the mature rRNAs (1,2). Interest in the nature of these processing sites has led investigators to sequence gene/spacer junctions, as well as the entire internal transcribed spacer regions in the cases of yeast, Xenopus, rat, and mouse (3-7). In yeast, the 3' termini of precursor and mature rRNAs share a sequence homology which suggests a recognition signal for processing; the same is true for the 5' terminus/spacer junctions (3,4). However, equivalent features are not apparent in Xenopus, rat, or mouse rDNA (5-7). Comparison of mature RNA-internal transcribed spacer junctions among species revealed no consensus sequences outside of the conserved termini of the rRNA (viz., 3 '-18S, 5'-5.8S, and 5'-26/28S), although some homologies were found between Xenopus and rat spacer regions (6).In this paper, we present the nucleotide sequence of a portion of

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Nucleotide sequence of the region between the 18S rRNA sequence and the 28S rRNA sequence of rat ribosomal DNA

Cited by 95 publications

References 24 publications

Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs.

Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs.

Biogenesis and nuclear export of ribosomal subunits in higher eukaryotes depend on the CRM1 export pathway

Nucleotide sequence of the 18S–26S rRNA intergene region of the sea urchin

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