Nucleic Acids Research, 1985, Vol. 13, No. 6 1871-1889
© 1985
Articles |
Sequence requirements for self-splicing of the Tetrahymena thermophila pre-ribosomal RNA
1Department of Molecular, Cellular and Developmental Biology Boulder, CO 80309, USA 2Department of Chemistry, University of Colorado Boulder, CO 80309, USA
Received January 18, 1985. Accepted February 14, 1985.
The sequence requirements for splicing of the Tetrahymena pre-rRNA have been examined by altering the rRNA gene to produce versions that contain insertions and deletions within the intervening sequence (IVS). The altered genes were transcribed and the RNA teeted for self-splicing in vitro. A number of insertions (8–54 nucleotides) at three locations had no effect on self-splicing activity. Two of these insertions, located at a site 5 nucleotides preceding the 3'-end of the IVS, did not alter the choice of the 3' splice site. Thus the 3' splice site is not chosen by its distance from a fixed point within the IVS. Analysis of deletions constructed at two sites revealed two structures, a hairpin loop and a stem-loop, that are entirely dispensable for IVS excision in vitro. Three other regions were found to be necessary. The regions that are important for self-splicing are not restricted to the conserved sequence element. that define this class of intervening sequences. The requirement for structures within the IVS for pre-rRNA splicing is in sharp contrast to the very limited role of IVS structure in nuclear pre-mRNA splicing.
+Present address: Amgen Development Inc., Boulder, CO 80301, USA
Present address: Baylor College of Medicine, Houston, TX 77030, USA
*Present address: Brown University, Providence, RI 02912, USA
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