Abstract
Using short oligoribonucleotides as ligated exon substrates, we show that splicing of the Tetrahymena rRNA group I intron is fully reversible in vitro. Incubation of ligated exon RNA with linear intron produces a molecule in which the splice site sequences of the precursor are reformed. Reversal of self-splicing is favored by high RNA concentration, high magnesium and temperature, and the absence of guanosine. 5′ exon sequences that can pair with the internal guide sequence of the intron are required, whereas 3′ exon sequences are not essential. Integration of the intron into ligated exon substrates that have the ability to form stem-loop structures is reduced at least one order of magnitude over short, unstructured substrates. We propose that the formation of these structures helps drive splicing in the forward direction. We also show that the Tetrahymena intron can integrate into a β-globin transcript. This has implications for transposition of group I introns.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 335-345 |
| Number of pages | 11 |
| Journal | Cell |
| Volume | 57 |
| Issue number | 2 |
| DOIs | |
| State | Published - Apr 21 1989 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
Cite this
Reverse self-splicing of the tetrahymena group I intron : Implication for the directionality of splicing and for intron transposition. / Woodson, Sarah A.; Cech, Thomas R.
In: Cell, Vol. 57, No. 2, 21.04.1989, p. 335-345.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Reverse self-splicing of the tetrahymena group I intron
T2 - Implication for the directionality of splicing and for intron transposition
AU - Woodson, Sarah A.
AU - Cech, Thomas R.
PY - 1989/4/21
Y1 - 1989/4/21
N2 - Using short oligoribonucleotides as ligated exon substrates, we show that splicing of the Tetrahymena rRNA group I intron is fully reversible in vitro. Incubation of ligated exon RNA with linear intron produces a molecule in which the splice site sequences of the precursor are reformed. Reversal of self-splicing is favored by high RNA concentration, high magnesium and temperature, and the absence of guanosine. 5′ exon sequences that can pair with the internal guide sequence of the intron are required, whereas 3′ exon sequences are not essential. Integration of the intron into ligated exon substrates that have the ability to form stem-loop structures is reduced at least one order of magnitude over short, unstructured substrates. We propose that the formation of these structures helps drive splicing in the forward direction. We also show that the Tetrahymena intron can integrate into a β-globin transcript. This has implications for transposition of group I introns.
AB - Using short oligoribonucleotides as ligated exon substrates, we show that splicing of the Tetrahymena rRNA group I intron is fully reversible in vitro. Incubation of ligated exon RNA with linear intron produces a molecule in which the splice site sequences of the precursor are reformed. Reversal of self-splicing is favored by high RNA concentration, high magnesium and temperature, and the absence of guanosine. 5′ exon sequences that can pair with the internal guide sequence of the intron are required, whereas 3′ exon sequences are not essential. Integration of the intron into ligated exon substrates that have the ability to form stem-loop structures is reduced at least one order of magnitude over short, unstructured substrates. We propose that the formation of these structures helps drive splicing in the forward direction. We also show that the Tetrahymena intron can integrate into a β-globin transcript. This has implications for transposition of group I introns.
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U2 - 10.1016/0092-8674(89)90971-9
DO - 10.1016/0092-8674(89)90971-9
M3 - Article
VL - 57
SP - 335
EP - 345
JO - Cell
JF - Cell
SN - 0092-8674
IS - 2
ER -