PMID: 8972845Dec 1, 1996Paper

In vitro reconstitution of mammalian U1 snRNPs active in splicing: the U1-C protein enhances the formation of early (E) spliceosomal complexes

Nucleic Acids Research
C L WillR Lührmann

Abstract

We have established an in vitro reconstitution/splicing complementation system which has allowed the investigation of the role of mammalian U1 snRNP components both in splicing and at the early stages of spliceosome formation. U1 snRNPs reconstituted from purified, native snRNP proteins and either authentic or in vitro transcribed U1 snRNA restored both early (E) splicing complex formation and splicing-activity to U1-depleted extracts. In vitro reconstituted U1 snRNPs possessing an m3G or ApppG cap were equally active in splicing, demonstrating that a physiological cap structure is not absolutely required for U1 function. However, the presence of an m7GpppG or GpppG cap was deleterious to splicing, most likely due to competition for the m7G cap binding proteins. No significant reduction in splicing or E complex formation was detected with U1 snRNPs reconstituted from U1 snRNA lacking the RNA binding sites of the U1-70K or U1-A protein (i.e., stem-loop I and II, respectively). Complementation studies with purified HeLa U1 snRNPs lacking subsets of the U1-specific proteins demonstrated a role for the U1-C, but not U1-A, protein in the formation and/or stabilization of early splicing complexes. Studies with recombinant U1-C protei...Continue Reading

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Citations

Jun 1, 1997·Current Opinion in Cell Biology·C L Will, R Lührmann
Dec 3, 2002·Insect Biochemistry and Molecular Biology·J M Sierra-MontesR J Herrera
Apr 5, 2012·Biochemistry·Clarisse van der FeltzDaniel A Pomeranz Krummel
Mar 28, 2009·Nature·Daniel A Pomeranz KrummelKiyoshi Nagai
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Dec 17, 2004·Molecular and Cellular Biology·Oliver A KentAndrew M Macmillan
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Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·M T BedfordP Leder
Oct 7, 2004·Proceedings of the National Academy of Sciences of the United States of America·Hansen DuMichael Rosbash
Jun 21, 2011·Biophysical Chemistry·Sandra G Williams, Kathleen B Hall
May 29, 2014·Biochemistry·Sandra G Williams, Kathleen B Hall
Oct 18, 2006·The FEBS Journal·Inês MolletMaria Carmo-Fonseca
Apr 7, 2011·The EMBO Journal·Tanja Dorothe RöselAlbrecht Bindereif
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