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| Status |
Public on Dec 31, 2009 |
| Title |
A systematic characterization of Cwc21, the yeast ortholog of the human spliceosomal protein SRm300 |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by genome tiling array
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| Summary |
Cwc21 (complexed with CEF1 protein 21) is a 135 amino acid yeast protein that shares homology with the N-terminal domain of human SRm300, a large serine/arginine-repeat protein shown previously to associate with the splicing coactivator 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.
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| Overall design |
S. cerevisiae mutants with single- and double-deletions for cwc21 and isy1 or cwc21 and bud13 were grown under standard conditions and genome-wide expression profiles were obtained using Affymetrix tiling microarrays. Pre-mRNA splicing defects were then assessed in pairwise comparisons with wild-type strains that were grown and assayed under the same conditions.
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| Contributor(s) |
Khanna M, van Bakel H, Tang Y, Calarco JA, Babak T, Guo G, Emili A, Greenblatt JF, Hughes TR, Krogan NJ, Blencowe BJ |
| Citation(s) |
19789211 |
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| Submission date |
Jul 29, 2009 |
| Last update date |
Mar 21, 2012 |
| Contact name |
Harm van Bakel |
| E-mail(s) |
harm.vanbakel@mssm.edu
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| Organization name |
Mount Sinai School of Medicine
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| Department |
Genetics and Genomic Sciences
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| Lab |
Bakel Lab
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| Street address |
One Gustave L. Levy Place, Box 1498
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| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10029 |
| Country |
USA |
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| Platforms (1) |
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| Samples (5)
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| GSM434588 |
cwc21-isy1-null expression profile |
| GSM434589 |
cwc21-bud13-null expression profile |
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| Relations |
| BioProject |
PRJNA118835 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE17395_RAW.tar |
422.8 Mb |
(http)(custom) |
TAR (of BAR, CEL) |
| Processed data provided as supplementary file |
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