Targeted snRNP depletion reveals an additional role for mammalian U1 snRNP in spliceosome assembly
CellPublished 1 October 1990
Silvia M.L. Barabino, Benjamin J. Blencowe, Ursula Ryder, Brian S. Sproat, Angus I. Lamond
Citations140
SJR quartileQ1
SJR score22.61
SNIP7.62
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TL;DR
HeLa cell nuclear splicing extracts have been prepared that are specifically and efficiently depleted of U1, U2, or U4/U6 snRNPs by antisense affinity chromatography using biotinylated 2'-OMe RNA oligonucleotides to promote stable preslicing complex formation.
Abstract
HeLa cell nuclear splicing extracts have been prepared that are specifically and efficiently depleted of U1, U2, or U4/U6 snRNPs by antisense affinity chromatography using biotinylated 2'-OMe RNA oligonucleotides. Removal of each snRNP particle prevents pre-mRNA splicing but arrests spliceosome formation at different stages of assembly. Mixing extracts depleted for different snRNP particles restores formation of functional splicing complexes. Specific binding of factors to the 3' splice site region is still detected in snRNP-depleted extracts. Depletion of U1 snRNP impairs stable binding of U2 snRNP to the pre-mRNA branch site. This role of U1 snRNP in promoting stable preslicing complex formation is independent of the U1 snRNA-5' splice site interaction.
Keywords
MedicineBiochemistry, Genetics and Molecular Biology
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