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Published Online October 31, 2002
Science DOI: 10.1126/science.1077783

Reports

Submitted on August 27, 2002
Accepted on October 21, 2002

Small Nuclear Ribonucleoprotein Remodeling During Catalytic Activation of the Spliceosome

Evgeny M. Makarov 1, Olga V. Makarova 1, Henning Urlaub 1, Marc Gentzel 2, Cindy L. Will 1, Matthias Wilm 2, Reinhard Lührmann 1*

1 Department of Cellular Biochemistry, Max Planck Institute of Biophysical Chemistry, D-37077 Göttingen, Germany.
2 EMBL, Bioanalytical Research Group, D-69117 Heidelberg, Germany.

* To whom correspondence should be addressed. E-mail: reinhard.luehrmann{at}mpi-bpc.mpg.de.

Major structural changes occur in the spliceosome during its activation just prior to catalyzing the splicing of pre-mRNAs. While changes in small nuclear RNA (snRNA) conformation are well documented, little is known about remodeling of small nuclear ribonucleoprotein (snRNP) structures during spliceosome activation. Here, human 45S activated spliceosomes and a novel 35S U5 snRNP were isolated by immunoaffinity selection and characterized by mass spectrometry. Comparison of their protein components with those of other snRNP and spliceosomal complexes revealed a major change in protein composition during spliceosome activation. Our data also suggest that the U5 snRNP is dramatically remodeled at this stage, with the Prp19 complex and other factors tightly associating, possibly in exchange for other U5 proteins, and that after catalysis the remodeled U5 is eventually released from the post-splicing complex as a 35S snRNP particle.


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