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Nucleic Acids Research, 1994, Vol. 22, No. 24 5190-5195
© 1994

Articles

The interaction between the first and last intron nucleotides in the second step of pre-mRNA splicing is independent of other conserved intron nucleotides

Brian L. Ruis, Wendy J. Kivens and Paul G. Siliciano*

Department of Biochemistry and Institute of Human Genetics, University of Minnesota Medical School 4-225 Millard Hall, 435 Delaware Street SE, Minneapolis, MN 55455, USA

*To whom correspondence should be addressed

Received October 4, 1994. Accepted October 28, 1994.

Virtually all pre-mRNA introns begin with the sequence / GU and end with AG/ (where / indicates a border between an exon and an Intron). We have previously shown that the G residues at the first and last positions of the yeast act In Intron Interact during the second step of splicing. In this work, we ask If other highly conserved Intron nucleotides also take part In this / G - G/ interaction. Of special interest is the penultimate Intron nucleotlde (AGl), which Is important for the second step of splicing and is In proximity to other conserved intron nucleotides. Therefore, we tested interactions of the penultimate intron nucleotide with the second intron nucleotide (/GU) and with the branch site nucleotide. We also tested two models that predict Interactions between sets of three conserved Intron nucleotides. In addition, we used random mutagenesls and genetic selection to search for Interactions between nucleotides In the pre-mRNA. We find no evidence for other Interactions between intron nucleotides besides the interaction between the first and last intron nucleotides.


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