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A systematic characterization of Cwc21, the yeast ortholog of the human spliceosomal protein SRm300.
- Source :
-
RNA (New York, N.Y.) [RNA] 2009 Dec; Vol. 15 (12), pp. 2174-85. Date of Electronic Publication: 2009 Sep 29. - Publication Year :
- 2009
-
Abstract
- Cwc21 (complexed with Cef1 protein 21) is a 135 amino acid yeast protein that shares homology with the N-terminal domain of human SRm300/SRRM2, a large serine/arginine-repeat protein shown previously to associate with the splicing coactivator and 3'-end processing stimulatory factor, SRm160. Proteomic analysis of spliceosomal complexes has suggested a role for Cwc21 and SRm300 at the core of the spliceosome. However, specific functions for these proteins have remained elusive. In this report, we employ quantitative genetic interaction mapping, mass spectrometry of tandem affinity-purified complexes, and microarray profiling to investigate genetic, physical, and functional interactions involving Cwc21. Combined data from these assays support multiple roles for Cwc21 in the formation and function of splicing complexes. Consistent with a role for Cwc21 at the core of the spliceosome, we observe strong genetic, physical, and functional interactions with Isy1, a protein previously implicated in the first catalytic step of splicing and splicing fidelity. Together, the results suggest multiple functions for Cwc21/SRm300 in the splicing process, including an important role in the activation of splicing in association with Isy1.
- Subjects :
- Carrier Proteins genetics
Humans
Oligonucleotide Array Sequence Analysis
Protein Binding
Proteomics
RNA, Small Nuclear genetics
RNA-Binding Proteins genetics
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins genetics
Carrier Proteins metabolism
RNA Splicing
RNA-Binding Proteins metabolism
Saccharomyces cerevisiae metabolism
Saccharomyces cerevisiae Proteins metabolism
Spliceosomes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1469-9001
- Volume :
- 15
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- RNA (New York, N.Y.)
- Publication Type :
- Academic Journal
- Accession number :
- 19789211
- Full Text :
- https://doi.org/10.1261/rna.1790509