Genetic Profiling of Protein Burden and Nuclear Export Overload

Abstract

Overproduction (op) of proteins triggers cellular defects. One of the defined consequences of protein overproduction is the protein burden/cost, which is produced by an overloading of the protein synthesis process. However, the physiology of cells under a protein burden is not well characterized. We performed genetic profiling of protein burden by systematic analysis of genetic interactions between GFP-op, surveying both deletion mutants of nonessential genes and temperature-sensitive mutants of essential genes, in the budding yeast Saccharomyces cerevisiae. To dissect interactions specific to the protein burden, we also performed genetic profiling in cells with overproduction of triple-GFP (tGFP), and the nuclear export signal-containing tGFP (NES-tGFP). The mutants specifically interacted with GFP-op were suggestive of unexpected connections between actin-related processes like polarization and the protein burden, which was supported by morphological analysis. The tGFP-op interactions suggested that this protein probe overloads the proteasome, probably through the formation of intracellular aggregates, whereas those that interacted with NES-tGFP involved genes encoding components of the nuclear export process, providing a resource for further analysis of the protein burden and nuclear export overload.