Reverse self-splicing of the tetrahymena group I intron

Implication for the directionality of splicing and for intron transposition

Sarah A. Woodson, Thomas R. Cech

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)335-345
Number of pages11
JournalCell
Volume57
Issue number2
DOIs
StatePublished - Apr 21 1989
Externally publishedYes

Fingerprint

Tetrahymena
Introns
Exons
Substrates
Oligoribonucleotides
RNA
Globins
Guanosine
Magnesium
Molecules
Temperature

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 journalArticle

@article{68399fdd06d84fe8a04c864f3d32b6bb,
title = "Reverse self-splicing of the tetrahymena group I intron: Implication for the directionality of splicing and for intron transposition",
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.",
author = "Woodson, {Sarah A.} and Cech, {Thomas R.}",
year = "1989",
month = "4",
day = "21",
doi = "10.1016/0092-8674(89)90971-9",
language = "English (US)",
volume = "57",
pages = "335--345",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "2",

}

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.

UR - http://www.scopus.com/inward/record.url?scp=0024975570&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024975570&partnerID=8YFLogxK

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 -