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Nucleic Acids Research, 1991, Vol. 19, No. 14 3857-3860
© 1991

MOLECULAR BIOLOGY

Genetic depletion indicates a late role for U5 snRNP during in vitro spliceosome assembly

Bertrand Séraphin*, Nadja Abovich and Michael Rosbash

Howard Hughes Medical Institute, Department of Biology, Brandeis University Waltham, MA 02254, USA

Received May 16, 1991. Revised June 18, 1991. Accepted June 18, 1991.

The pre-mRNA splicing pathway is highly conserved from yeast (S. cerevisiae) to mammals. Of the four snRNPs involved in splicing three (U1, U2 and U4/U6) have been shown to be essential for in vitro splicing. To examine the remaining snRNP, we utilized our previously described genetic procedures (Seraphin and Rosbash, 1989) to prepare yeast extracts depleted of U5 snRNP. The results show that U5 snRNP is necessary for both steps of pre- mRNA splicing and for proper spllceosome assembly, i.e., addition of the U4/U5/U6 triple snRNP. The prior steps of U1 and U2 snRNP addition occur normally in the absence of U5 snRNP.

* Present address: European Molecular Biology Laboratory, Postfach 10.2209, Meyerhofstrasse 1, D-6900 Heidelberg, FRG


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