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. 1988 Oct 25;16(20):9415-29.
doi: 10.1093/nar/16.20.9415.

Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 and U5 snRNPs

A R Krainer  1
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Free PMC article

Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 and U5 snRNPs

A R Krainer. Nucleic Acids Res. .
Free PMC article

Abstract

The four major nucleoplasmic small nuclear ribonucleoprotein particles U1, U2, U4/U6 and U5 can be extensively purified from HeLa cells by immunoaffinity chromatography using a monoclonal anti-trimethylguanosine antibody. The snRNP particles in active splicing extracts are selectively bound to the immunoaffinity matrix, and are then gently eluted by competition with an excess of free nucleoside. Biochemical complementation studies show that the purified snRNPs are active in pre-mRNA splicing, but only in the presence of additional non-snRNP protein factors. All the RNPs that are necessary for splicing can be purified in this manner. The active snRNPs are characterized with respect to their polypeptide composition, and shown to be distinct from several other activities implicated in splicing.

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