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Partial purification of the yeast U2 snRNP reveals a novel yeast pre‐mRNA splicing factor required for pre‐spliceosome assembly
Author(s) -
Caspary Friederike,
Shevchenko Anna,
Wilm Matthias,
Séraphin Bertrand
Publication year - 1999
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/18.12.3463
Subject(s) - biology , snrnp , spliceosome , rna splicing , yeast , splicing factor , genetics , precursor mrna , saccharomyces cerevisiae , small nuclear rna , intron , computational biology , rna , gene , non coding rna
We have partially purified the U2 snRNP of Saccharomyces cerevisiae . Identification of some proteins consistently found in the purified fractions by nanoelectrospray mass spectrometry indicated the presence of a novel splicing factor named Rse1p. The RSE1 gene is essential and codes for a 148.2 kDa protein. We demonstrated that Rse1p associates specifically with U2 snRNA at low salt concentrations. In addition, we showed that Rse1p is a component of the pre‐spliceosome. Depletion of Rse1p and analysis of a conditional mutant indicated that Rse1p was required for efficient splicing in vivo . In vitro Rse1p is required for the formation of pre‐spliceosomes. Database searches revealed that Rse1p is conserved in humans and that it belongs to a large protein family that includes polyadenylation factors and DNA repair proteins. The characteristics of Rse1p suggest that its human homologue could be a subunit of the SF3 splicing factor.