Isolation of S. cerevisiae snRNPs: comparison of U1 and U4/U6.U5 to their human counterparts.
Small nuclear ribonucleoprotein (snRNP) particles are essential for pre-messenger RNA splicing. In human HeLa cells, 40 proteins associated with snRNPs have been identified. Yet, the function of many of these proteins remains unknown. Here, the immunoaffinity purification of the spliceosomal snRNPs U1, U2, U4/U6.U5, and several nucleolar snRNP species from the yeast Saccharomyces cerevisiae is presented. The U1 and U4/U6.U5 snRNPs were purified extensively and their protein composition and ultrastructure analyzed. The yeast U1 snRNP is larger and contains three times more specific proteins than its human counterpart. In contrast, the size, protein composition, and morphology of the yeast and the human U4/U6.U5 snRNPs are significantly similar. The preparative isolation of yeast snRNPs will allow the cloning as well as genetic and phylogenetic analysis of snRNP proteins which will accelerate our understanding of their function.
Fabrizio
P
P
Institut für Molekularbiologie und Tumorforschung, Phillipps-Universität, Marburg, Germany.
Esser
S
S
Kastner
B
B
Lührmann
R
R
D015153:Blotting, Western; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D008970:Molecular Weight; D012331:RNA, Fungal / Q000032:analysis; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000737:chemistry / Q000302:isolation & purification; D017414:Ribonucleoprotein, U4-U6 Small Nuclear / Q000737:chemistry / Q000302:isolation & purification; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000737:chemistry / Q000302:isolation & purification; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D017381:Spliceosomes / Q000737:chemistry*
D012331:RNA, Fungal; D017412:Ribonucleoprotein, U1 Small Nuclear; D017414:Ribonucleoprotein, U4-U6 Small Nuclear; D017415:Ribonucleoprotein, U5 Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear