Global proteomics proteasome mutants

ABSTRACT: Temperature sensitive(TS)missense mutants have been foundational in the characterization of the function of essential genes due to their pronounced cellular phenotypes that are likely associated with protein functional disruption. However, an unbiased approach for the analysis of the biochemical and biophysical changes in missense mutants within the context of their functional proteomes is lacking. We applied mass spectrometry (MS) based thermal proteome profiling (TPP)to investigate the proteome-wide effects of missense mutations in a specific application of TPP that we refer to as mutant Thermal Proteome Profiling (mTPP). 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 mTPP 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Sarah Peck Justice  

LAB HEAD: Amber L Mosley

PROVIDER: PXD017223 | Pride | 2020-11-03

REPOSITORIES: pride