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| Status |
Public on Jan 08, 2020 |
| Title |
Temperature sensitive Mutant Proteome Profiling: a novel tool for the characterization of the global impacts of missense mutants on the proteome |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
RNA-seq experiments measuring global RNA abundance
Temperature sensitive (TS) mutants are a tool that have been foundational for the study of many essential life processes. Despite the long-term use of TS mutants, the mechanisms that lead to temperature sensitivity are not fully understood. Furthermore, a high-throughput workflow to characterize biophysical changes occurring in TS mutants is lacking. We developed Temperature sensitive Mutant Proteome Profiling (TeMPP), a novel application of mass spectrometry based thermal proteome profiling (TPP) as a way to measure the effects of missense mutations on protein stability and protein-protein interactions. This study characterized the global changes in mRNA abundance, protein abundance, and protein thermal stability as a result of missense mutants within two subunits of the yeast ubiquitin-proteasome system. Global protein abundance measurements and RNA sequencing data resulted in a large number of possible candidates that could be causing the phenotypic changes observed in the mutant strains. The additional information gained from TeMPP along with complementary proteomic and transcriptomic experiments allows for multiomic intersection analysis that may reveal interesting regulatory categories to pursue in follow-up mechanistic experiments.
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| Overall design |
Four RNA-Seq biological replicates each for ts mutant (pup2-ts and rpn5-ts) and reference cells (WT)
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| |
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| Contributor(s) |
Justice SA, Qi G, Wijeratne HS, Victorino JF, Simpson ER, Wijeratne AB, Mosley AL |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Jan 07, 2020 |
| Last update date |
Jan 08, 2020 |
| Contact name |
Sarah Peck Justice |
| E-mail(s) |
sapeck@iupui.edu
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| Organization name |
Indiana University
|
| Department |
Biochemistry and Molecular Biology
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| Lab |
Mosley
|
| Street address |
635 Barnhill Dr
|
| City |
Indianapolis |
| State/province |
IN |
| ZIP/Postal code |
46220 |
| Country |
USA |
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| Platforms (1) |
| GPL21656 |
Illumina HiSeq 4000 (Saccharomyces cerevisiae) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA599371 |
| SRA |
SRP239803 |
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