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Science 24 December 1993:
Vol. 262. no. 5142, pp. 1982 - 1988
DOI: 10.1126/science.8266093
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Articles

Science, Vol 262, Issue 5142, 1982-1988
Copyright © 1993 by American Association for the Advancement of Science

articles

Mutations in U6 snRNA that alter splice site specificity: implications for the active site

CF Lesser and C Guthrie

Medical Scientist Training Program, University of California, San Francisco 94143.

What determines the precise sites of cleavage in the two transesterification reactions of messenger RNA (mRNA) splicing is a major unsolved question. Mutation of the invariant G (guanosine) at position 5 of 5' splice sites in Saccharomyces cerevisiae introns activates cleavage at nearby aberrant sites. A genetic approach was used to test the hypothesis that a base-pairing interaction between the 5' splice site and the invariant ACAGAG sequence of U6 is a determinant of 5' splice site choice. Mutations in U6 or the intron (or both) that were predicted to stabilize the interaction suppressed aberrant cleavage and increased normal cleavage. In addition, a mutation in the ACAGAG sequence suppressed mutations of the 3' splice site dinucleotide. These data can fit a model for the spliceosomal active site comprised of a set of RNA-RNA interactions between the intron, U2 and U6.


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Guides to the heart of the spliceosome.
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The U5 and U6 small nuclear RNAs as active site components of the spliceosome.
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Involvement of U6 snRNA in 5' splice site selection.
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