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Science 23 September 1994:
Vol. 265. no. 5180, pp. 1866 - 1869
DOI: 10.1126/science.8091213
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Articles

Science, Vol 265, Issue 5180, 1866-1869
Copyright © 1994 by American Association for the Advancement of Science

articles

Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP

JD Crispino, BJ Blencowe, and PA Sharp

Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.

Individual small nuclear ribonucleoproteins (snRNPs) U1, U2, and U4/U6 were removed from nuclear extracts of HeLa cells by antisense affinity depletion. Addition of a highly purified preparation of SR proteins fully restored splicing activity in reactions depleted of U1 snRNP but did not reconstitute splicing in reactions depleted of the other snRNPs. Affinity selection experiments revealed that spliceosomes lacking U1 snRNA formed in the U1 snRNP-depleted reactions reconstituted with SR proteins. Thus, high concentrations of SR proteins facilitate the assembly of precursor messenger RNA (pre-mRNA) into a spliceosome in the absence of interactions with U1 snRNP.


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