The DOI matches its Crossref record; 38 of the 39 references it cites were found in a registry or an index.
In vitro splicing of the ribosomal RNA precursor in nuclei of tetrahymena
Abstract
The macronuclear rRNA genes of Tetrahymena thermophila contain a 0.4 kb intervening sequence. In this paper we present evidence that the excision of the intervening sequence from pre-rRNA occurs in vitro in isolated T. thermophila nuclei. The transcription-processing system includes alpha-amanitin to inhibit non-rRNA synthesis and aurintricarboxylic acid to inhibit endogenous nucleases. A discrete 0.4 kb RNA comprises up to 6% of the RNA synthesized in this system. Southern hybridization with restriction fragments of the rDNA (rRNA genes) shows that the 0.4 kb RNA contains the intervening sequence. The size of the 0.4 kb RNA, 410-425 nucleotides, is the size predicted if the entire intervening sequence were excised as a single linear molecule. The 0.4 kb RNA accumulates post-transcriptionally and appears to be stable in vitro. T. pigmentosa strain 6UM, whose rDNA has an intervening sequence of the same size and location as that of T. thermophila, also produces a 0.4 kb RNA in vitro; T. pigmentosa strain 8ALP, with not rDNA intervening sequence, produces no such RNA. The measurement of the accumulation of the excised intervening sequence is a convenient assay for the pre-rRNA splicing activity and a means for characterizing some of the details of the reaction. The resistance of the splicing acticity to concentrations of aurintricarboxylic acid that inhibit other endogenous nucleases should be useful in assaying the splicing enzyme during purification.
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“In 1982, this conflict was resolved by the discovery of ribozymes, catalytic RNA molecules [114,115].”
From Opinion: The Key Steps in the Origin of Life to the Formation of the Eukaryotic Cell (2024) · cites this paper as [114]
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Cites
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RNA Processing and the Intervening Sequence Problem
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The structural organization of ribosomal DNA in drosophila melanogaster
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Messenger RNA in heat-shocked Drosophila cells
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Use of aurintricarboxylic acid as an inhibitor of nucleases during nucleic acid isolation
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A novel arrangement of the 18S and 28S sequences in a repeating unit of drosophila melanogaster rDNA
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Metadata
- Authors
- Zaug, A. J., & Cech, T. R.
- Affiliations
- University of Colorado Boulder (US)
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 1980-02
- Volume
- 19
- Issue
- 2
- Pages
- 331-338
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2046737757
- Cited by
- 115
- References
- 39
- Citations by year
- 2026: 1 · 2024: 2 · 2023: 4 · 2022: 6 · 2021: 2 · 2019: 1 · 2018: 3 · 2017: 3 · 2016: 2 · 2015: 2 · 2014: 1
- FWCI
- 5.14
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- ribosomal precursor RNA, Tetrahymena thermophila, post-transcriptional processing, aurintricarboxylic acid, alpha-amanitin
- MeSH terms
- Animals, Base Sequence, Cell Nucleus, DNA, Nucleic Acid Hybridization, Nucleic Acid Precursors, RNA, Ribosomal, Tetrahymena, Transcription, Genetic
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