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PubMed · 1835087

Two-step affinity purification of U7 small nuclear ribonucleoprotein particles using complementary biotinylated 2'-O-methyl oligoribonucleotides.

H O Smith·K Tabiti·G Schaffner·D Soldati·U Albrecht·M L Birnstiel
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Abstract

U7 snRNP is a low-abundance small nuclear ribonucleoprotein particle essential for 3' processing of replication-dependent histone pre-mRNA. We have developed a two-step purification of the particle from TB21 mouse mastocytoma cell nuclear extracts, with about a 20% overall yield, using affinity binding to 2'-O-methyl oligoribonucleotides. The purified particle is homogeneous with respect to RNA content. SDS/PAGE of the U7 snRNP proteins revealed a full complement of the standard core proteins (B, DD', E, F, and G) found in the majority of snRNPs. In addition, two U7-specific polypeptides of 14 kDa and 50 kDa were identified. Summation of the molecular masses of the identified components of the U7 particle yields a particle mass of 249 kDa, in approximate agreement with estimates from sucrose gradient sedimentation (261 kDa) and nondenaturing gradient PAGE (217 kDa).

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AnimalsBase SequenceBiotinCell NucleusChromatography, AffinityMiceMolecular Sequence DataMolecular WeightNucleic Acid HybridizationOligonucleotidesRibonucleoproteinsRibonucleoproteins, Small NuclearJournal ArticleResearch Support, Non-U.S. Gov't

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H O Smith, K Tabiti, G Schaffner, D Soldati, U Albrecht, M L Birnstiel. 1991-11-01. Two-step affinity purification of U7 small nuclear ribonucleoprotein particles using complementary biotinylated 2'-O-methyl oligoribonucleotides.. https://doi.org/10.1073/pnas.88.21.9784

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