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Nucleic Acids Research, 1985, Vol. 13, No. 23 8389-8408
© 1985

Articles

Sites of circularization of the Tetrahymena rRNA IVS are determined by sequence and influenced by position and secondary structure

Michael D. Been and Thomas R. Cech

Department of Chemistry and Biochemistry, University of Colorado Boulder, CO 80309, USA

Received September 23, 1985. Accepted November 4, 1985.

The sequence of the cloned Tetrahymena ribosomal RNA intervening sequence (IVS) was altered at the site to which circularization normally occurs. The alterations caused circularization to shift to other sites, usually a nearby position which followed three pyrimidines. While a tripyrimidine sequence was the major determinant of a circularization site, both location of a sequence and local secondary structure may influence the use of that sequence. For some constructs circularization appeared to occur at the position following the 5' G, the nucleotide added to the IVS during its excision. Portions of the internal guide sequence (IGS), proposed to interact with the 3'exon were deleted without preventing exon ligation. Thus if the IGS-3'exon interaction exists, it is not essential for splicing in vitro.


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