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Nucleic Acids Research, 1989, Vol. 17, No. 1 335-354
© 1989

MOLECULAR BIOLOGY

Sequence requirements for branch formation in a group ll self-splicing lntron

Rachel Altura+, Brian Rymond§, Bertrand Seraphin and Michael Rosbash

Department of Biology, Brandeis University Waltham, MA 02254, USA

Received August 5, 1988. Revised November 28, 1988. Accepted November 28, 1988.

Evidence is presented for the existence of a specific intronintron interaction, necessary for the formation of the branched product in the self-splicing reaction of a group II yeast mitochondrial intron. Trans-splicing reactions involving two RNA molecules (5'exon with covalently linked regions of intron and intron with covalently linked 3'exon) show that the presence of portions of intron domain I on the 51 molecule is necessary for the formation of branched products which are not seen with shorter 51 molecules. Modification/interference reactions show regions necessary for branch-formation and support a major role for specific regions of intron domain I. Further experiments, utilizing a truncated 31 molecule that is missing the conserved branchpoint nucleotide, indicate that domain VI may be required for a successful domain I interaction. A model for the formation of a proper branched structure includes implications for both cis and trans configurations.

+Present addresses: Washington University, School of Medicine, St Louis, MO 63110

§Present addresses: University of Kentucky, School of Biological Sciences, Lexington, KY 40506-0225, USA


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