Self-splicing RNA: Autoexcision and autocyclization of the ribosomal RNA intervening sequence of tetrahymena

@article{Kruger1982SelfsplicingRA,
  title={Self-splicing RNA: Autoexcision and autocyclization of the ribosomal RNA intervening sequence of tetrahymena},
  author={K E Kruger and Paula J. Grabowski and Arthur J. Zaug and Julie Sands and Daniel E. Gottschling and Thomas R. Cech},
  journal={Cell},
  year={1982},
  volume={31},
  pages={147-157}
}
In the macronuclear rRNA genes of Tetrahymena thermophila, a 413 bp intervening sequence (IVS) interrupts the 26S rRNA-coding region. A restriction fragment of the rDNA containing the IVS and portions of the adjacent rRNA sequences (exons) was inserted downstream from the lac UV5 promoter in a recombinant plasmid. Transcription of this template by purified Escherichia coli RNA polymerase in vitro produced a shortened version of the pre-rRNA, which was then deproteinized. When incubated with… Expand
Fate of an intervening sequence ribonucleic acid: excision and cyclization of the Tetrahymena ribosomal ribonucleic acid intervening sequence in vivo.
TLDR
It is concluded that splicing is not a rate-limiting step in rRNA maturation and that the IVS RNA is quickly degraded after its excision, and Autocyclization, previously characterized withIVS RNA produced by splicing in vitro, is therefore also a property of IVSRNA produced in vivo. Expand
Mechanism and requirements of in vitro RNA splicing of the primary transcript from the T4 bacteriophage thymidylate synthase gene.
TLDR
The splicing of a procaryotic precursor RNA transcribed from the T4 phage thymidylate synthase (td) gene with SP6 RNA polymerase was investigated in vitro and the identification of various structural elements that are found typically in eucaryotic class I introns firmly establish the td intron as the first example of class I intron of Procaryotic origin. Expand
5' exon requirement for self-splicing of the Tetrahymena thermophila pre-ribosomal RNA and identification of a cryptic 5' splice site in the 3' exon.
TLDR
The intervening sequence (IVS) of the Tetrahymena thermophila ribosomal RNA precursor undergoes accurate self-splicing in vitro and predicts the existence of a circular RNA containing the first 52 nucleotides of the 3' exon, which can function as a cryptic 5' splice site. Expand
New reactions of the ribosomal RNA precursor of Tetrahymena and the mechanism of self-splicing.
TLDR
The finding that the splice site can act as a cyclized site, combined with the earlier observation that the normal cyclization site is subject to attack by guanosine mononucleotide, leads us to propose that all these reactions may be occurring in the same active site. Expand
Origin of the phosphate at the ligation junction produced by self-splicing of Tetrahymena thermophila pre-ribosomal RNA.
  • J. V. Price
  • Biology, Medicine
  • Journal of molecular biology
  • 1987
TLDR
It is demonstrated that the phosphate group that forms the phosphodiester bond at the ligation junction is derived from the 5' position of a uridine nucleotide in the RNA, which supports the prediction that the splice junction phosphate isderived from the 3' splice site. Expand
Conformational Dynamics Involved in RNA Self-Splicing
TLDR
In some species of the ciliated protozoan, Tetrahymena, the genes for the large rRNA are interrupted by an IVS, which is then subject to an RNA processing reaction called RNA splicing, by which the IVS is excised and the exons or flanking sequences are ligated. Expand
Intermolecular exon ligation of the rRNA precursor of tetrahymena: Oligonucleotides can function as 5′ exons
TLDR
The mechanism by which 5' splice sites are selected in Tetrahymena rRNA and group I mitochondrial RNA splicing is like that used in nuclear mRNA splicing, in that it involves specific pairing of bases adjacent to the splice site with a complementary RNA sequence. Expand
R2 Retrotransposons Encode a Self-Cleaving Ribozyme for Processing from an rRNA Cotranscript
TLDR
It is shown here that processing of the rRNA cotranscript at the 5′ end of the R2 element in Drosophila simulans is rapid and utilizes an unexpected mechanism, capable of rapid and efficient self-cleavage of the 28S-R2 cOTranscript. Expand
Reversibility of cyclization of the tetrahymena rRNA intervening sequence: implication for the mechanism of splice site choice
TLDR
It is found that certain di- and trinucleotides with free 3' hydroxyl groups reopen the circular IVS at the cyclization junction, producing a linear molecule with the oligonucleotide covalently attached to its 5' end. Expand
Self-splicing RNA: implications for evolution.
  • T. Cech
  • Biology, Medicine
  • International review of cytology
  • 1985
TLDR
This chapter discusses the evolution of self-splicing RNA and highlights Tetrahymena pre-rRNA splicing is self-catalyzed, which lowers the activation energy for a chemical reaction in which that same molecule is a reactant. Expand
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References

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In vitro splicing of the ribosomal RNA precursor in nuclei of tetrahymena
TLDR
Evidence that the excision of the intervening sequence from pre-rRNA occurs in vitro in isolated T. thermophila nuclei is presented, and the resistance of the splicing acticity to concentrations of aurintricarboxylic acid that inhibit other endogenous nucleases should be useful in assaying thesplicing enzyme during purification. Expand
In vitro splicing of the ribosomal RNA precursor of tetrahymena: Involvement of a guanosine nucleotide in the excision of the intervening sequence
TLDR
It is proposed that Tetrahymena pre-rRNA splicing occurs by a phosphoester transferase mechanism where the guanosine cofactor provides the free 3' hydroxyl necessary to initiate a series of three transfers that results in splicing of the pre- rRNA and cyclization of the excised IVS. Expand
The intervening sequence of the ribosomal RNA precursor is converted to a circular RNA in isolated nuclei of tetrahymena
TLDR
The finding of a novel RNA molecule, which is an electrophoretic variant (EV) of this 0.4 kb IVS RNA, which survived treatment with DNAase, protease, glyoxal and various denaturants, which suggests that it is a covalently closed RNA circle. Expand
In vitro splicing of the ribosomal RNA precursor in isolated nucleoli from Tetrahymena.
TLDR
It is demonstrated that the enzyme activities necessary for the endonucleolytic cleavage as well as for the ligation of the transcript are associated with the isolated macronuclear rRNA genes of Tetrahymena thermophila. Expand
Localization of transcribed regions on extrachromosomal ribosomal RNA genes of Tetrahymena thermophila by R-loop mapping.
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  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1979
TLDR
R-loop hybridization and electron microscopy were used to map the RNA transcription products of the extrachromosomal rRNA genes of Tetrahymena thermophila, and preliminary evidence for a primary transcript that contains the intervening sequence was found. Expand
The intervening sequence excised from the ribosomal RNA precursor of Tetrahymena contains a 5-terminal guanosine residue not encoded by the DNA.
TLDR
The splicing mechanisms are fundamentally different for yeast tRNA intervening sequences and Tetrahymena thermophila transcripts, leading to the conclusion that the splicing mechanism for these two classes of transcripts is fundamentally different. Expand
The nucleotide sequence at the transcription termination site of the ribosomal RNA gene in Tetrahymena thermophila.
TLDR
The sequence of 415 nucleotides surrounding the transcription termination site for ribosomal RNA in Tetrahymena thermophila has been determined and a cluster of T's at the putative termination site, and several other T clusters are found further downstream. Expand
The intervening sequence in the 26s rrna coding region of t. thermophila is transcribed within the largest stable precursor for rRNA
TLDR
It is concluded that the intervening sequence is transcribed within the primary transcription product of the rDNA, and that the post-transcriptional removal of the intervening RNA sequence is a very early processing event in the organism. Expand
Detailed transcription map of the extrachromosomal ribosomal RNA genes in Tetrahymena thermophila.
TLDR
It is shown that Tetrahymena exemplifies a case of a lower eukaryote in which the loss of transcribed spacer material during the processing of the pre-rRNA is very low (less than 20%). Expand
Sequence of a ribosomal RNA gene intron from Tetrahymena
TLDR
The sequence of a ribosomal RNA gene intron from Tetrahymena is reported in which intron–exon junctions differ from those analysed to date, suggesting different splicing activities are necessary for the processing of introns in mRN A and tRN A precursors. Expand
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