Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA

Roy Parker; Paul G. Siliciano; Christine Guthrie

doi:10.1016/0092-8674(87)90564-2

Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA

Roy Parker
, Paul G. Siliciano
, Christine Guthrie

Administration (OUE)

Research output: Contribution to journal › Article › peer-review

365 Scopus citations

Abstract

The U2 snRNP binds to the site of branch formation during splicing of mammalian pre-mRNA in vitro. In Saccharomyces cerevisiae the branch site is within the so-called TACTAAC box (UACUAAC box), an absolutely conserved intron sequence required for splicing. Based on the identification and sequence of a U2 analogue in yeast, a specific base pairing interaction between the UACUAAC box and a highly conserved region of this snRNA can be proposed. To test this hypothesis, we have taken advantage of two mutations constructed previously in the UACUAAC box of an actin-HIS4 fusion. These mutant strains were transformed with stable plasmids bearing U2-like snRNAs into which changes predicted to restore base pairing had been introduced. Aliele-specific suppression of biological and biochemical phenotypes was observed in both cases. Recognition of the UACUAAC box thus relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue.

Original language	English (US)
Pages (from-to)	229-239
Number of pages	11
Journal	Cell
Volume	49
Issue number	2
DOIs	https://doi.org/10.1016/0092-8674(87)90564-2
State	Published - Apr 24 1987

Bibliographical note

Funding Information:
We are grateful to M. Ares for communicating results prior to publication. We thank L. Esperas for excellent technical assistance, and J. Pic-cini for help in preparation of the manuscript. This work was supported by grants GM21119 from the National Institutes of Health and DCB-6603926 from the National Science Foundation to C. G. R G. S. was supported by a Damon Runyon-Walter Winchell Cancer Fund Fellowship (DRG-672).

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title = "Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA",

abstract = "The U2 snRNP binds to the site of branch formation during splicing of mammalian pre-mRNA in vitro. In Saccharomyces cerevisiae the branch site is within the so-called TACTAAC box (UACUAAC box), an absolutely conserved intron sequence required for splicing. Based on the identification and sequence of a U2 analogue in yeast, a specific base pairing interaction between the UACUAAC box and a highly conserved region of this snRNA can be proposed. To test this hypothesis, we have taken advantage of two mutations constructed previously in the UACUAAC box of an actin-HIS4 fusion. These mutant strains were transformed with stable plasmids bearing U2-like snRNAs into which changes predicted to restore base pairing had been introduced. Aliele-specific suppression of biological and biochemical phenotypes was observed in both cases. Recognition of the UACUAAC box thus relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue.",

author = "Roy Parker and Siliciano, \{Paul G.\} and Christine Guthrie",

note = "Funding Information: We are grateful to M. Ares for communicating results prior to publication. We thank L. Esperas for excellent technical assistance, and J. Pic-cini for help in preparation of the manuscript. This work was supported by grants GM21119 from the National Institutes of Health and DCB-6603926 from the National Science Foundation to C. G. R G. S. was supported by a Damon Runyon-Walter Winchell Cancer Fund Fellowship (DRG-672).",

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T1 - Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA

AU - Parker, Roy

AU - Siliciano, Paul G.

AU - Guthrie, Christine

N1 - Funding Information: We are grateful to M. Ares for communicating results prior to publication. We thank L. Esperas for excellent technical assistance, and J. Pic-cini for help in preparation of the manuscript. This work was supported by grants GM21119 from the National Institutes of Health and DCB-6603926 from the National Science Foundation to C. G. R G. S. was supported by a Damon Runyon-Walter Winchell Cancer Fund Fellowship (DRG-672).

PY - 1987/4/24

Y1 - 1987/4/24

N2 - The U2 snRNP binds to the site of branch formation during splicing of mammalian pre-mRNA in vitro. In Saccharomyces cerevisiae the branch site is within the so-called TACTAAC box (UACUAAC box), an absolutely conserved intron sequence required for splicing. Based on the identification and sequence of a U2 analogue in yeast, a specific base pairing interaction between the UACUAAC box and a highly conserved region of this snRNA can be proposed. To test this hypothesis, we have taken advantage of two mutations constructed previously in the UACUAAC box of an actin-HIS4 fusion. These mutant strains were transformed with stable plasmids bearing U2-like snRNAs into which changes predicted to restore base pairing had been introduced. Aliele-specific suppression of biological and biochemical phenotypes was observed in both cases. Recognition of the UACUAAC box thus relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue.

AB - The U2 snRNP binds to the site of branch formation during splicing of mammalian pre-mRNA in vitro. In Saccharomyces cerevisiae the branch site is within the so-called TACTAAC box (UACUAAC box), an absolutely conserved intron sequence required for splicing. Based on the identification and sequence of a U2 analogue in yeast, a specific base pairing interaction between the UACUAAC box and a highly conserved region of this snRNA can be proposed. To test this hypothesis, we have taken advantage of two mutations constructed previously in the UACUAAC box of an actin-HIS4 fusion. These mutant strains were transformed with stable plasmids bearing U2-like snRNAs into which changes predicted to restore base pairing had been introduced. Aliele-specific suppression of biological and biochemical phenotypes was observed in both cases. Recognition of the UACUAAC box thus relies, at least in part, on Watson-Crick base pairing with the yeast U2 analogue.

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Recognition of the TACTAAC box during mRNA splicing in yeast involves base pairing to the U2-like snRNA

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