Isolation of U3 snoRNP from CHO cells: a novel 55 kDa protein binds to the central part of U3 snoRNA
| Autor: | Christopher Marshallsay, Norbert Rottmann, Birgit Lübben, Reinhard Lührmann |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Fibrillarin
Binding Sites Base Sequence Nucleolus Molecular Sequence Data Nuclear Proteins Plasma protein binding CHO Cells Biology Ribonucleoproteins Small Nuclear Molecular biology Molecular Weight Structure-Activity Relationship Biochemistry Cricetinae RNA Small Nuclear Genetics Animals Nucleic Acid Conformation Small nucleolar RNA Nuclear protein Binding site Sequence motif Cell Nucleolus Ribonucleoprotein |
| Zdroj: | Nucleic acids research. 21(23) |
| ISSN: | 0305-1048 |
| Popis: | U3 snoRNP, the most abundant of the small nucleolar ribonucleoprotein particles (snoRNPs), has previously been demonstrated to participate in pre-rRNA maturation. Here we report the purification of U3 snoRNP from CHO cells using anti-m3G-immunoaffinity and mono Q anion-exchange chromatography. Isolated U3 snoRNPs contain three novel proteins, of 15, 50 and 55 kDa respectively. These proteins may represent core U3 snoRNP proteins whose binding mediates the association of other proteins, such as fibrillarin, that are lost during purification. Using a rabbit antiserum raised against the 55 kDa protein, and an in vitro reconstitution assay, we have localised the 55 kDa protein binding site on the U3 snoRNA. Stable binding of the 55 kDa protein requires sequences located between nucleotides 97 and 204 of the human U3 snoRNA, including the evolutionarily conserved B and C sequence motifs. |
| Databáze: | OpenAIRE |
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