Systemic control of protein synthesis through sequestration of translation and ribosome biogenesis factors during severe heat stress.

Environmental stress causes the sequestration of proteins into insoluble deposits including cytoplasmic stress granules (SGs), containing mRNA and a variety of translation factors. Here we systematically identified proteins sequestered in Saccharomyces cerevisiae at 46 °C by a SG co-localization screen and proteomic analysis of insoluble protein fractions. We identified novel SG components including essential aminoacyl-tRNA synthetases. Moreover, we discovered nucleus-associated deposits containing ribosome biogenesis factors. Our study suggests downregulation of cytosolic protein synthesis and nuclear ribosome production at multiple levels through heat shock induced protein sequestrations.

Coordination of translational control and protein homeostasis during severe heat stress.

BACKGROUND: Exposure of cells to severe heat stress causes not only misfolding and aggregation of proteins but also inhibition of translation and storage of mRNA in cytosolic heat stress granules (heat-SGs), limiting newly synthesized protein influx into overloaded proteome repair systems. How these two heat stress responses connect is unclear. RESULTS: Here, we show that both S. cerevisiae and D. melanogaster heat-SGs contain mRNA, translation machinery components (excluding ribosomes), and molecular chaperones and that heat-SGs coassemble with aggregates of misfolded, heat-labile proteins. Components in these mixed assemblies exhibit distinct molecular motilities reflecting differential trapping. We demonstrate that heat-SG disassembly and restoration of translation activity during heat stress recovery is intimately linked to disaggregation of damaged proteins present in the mixed assemblies and requires Hsp104 and Hsp70 activity. CONCLUSIONS: Chaperone-driven protein disaggregation directly coordinates timing of translation reinitiation with protein folding capacity during cellular protein quality surveillance, enabling efficient protein homeostasis.

Expression pattern of human glutaredoxin 2 isoforms: identification and characterization of two testis/cancer cell-specific isoforms.

The cellular redox state is associated with major cellular processes including differentiation, transformation, and apoptosis. Glutaredoxin 2 (Grx2) is a mitochondrial oxidoreductase suggested to play a critical role in protection against apoptotic stimuli. An alternative Grx2 transcript variant encoding a nonmitochondrial protein (Grx2b) was proposed before, but no data was available on the expression of this isoform. We have systematically investigated the expression of Grx2 transcript variants in human tissues and transformed cell lines. The transcript variant encoding mitochondrial Grx2 (Grx2a) was found to be ubiquitously expressed, emphasizing the general importance of the protein for mitochondrial redox homeostasis. In addition, we confirmed the previously suggested isoform Grx2b and identified a new third isoform (Grx2c) derived from alternative splicing of the Grx2b-encoding transcript. In normal tissue expression of both Grx2b and Grx2c was restricted to testes, but additionally we were able to demonstrate transcripts in various cancer cell lines. Both Grx2b and Grx2c are enzymatically active, but only Grx2c can complex the regulatory iron-sulfur cluster described for Grx2a. Expression of GFP fusion proteins suggested a cytosolic and nuclear localization of both Grx2b and Grx2c. Our findings provide the first evidence for functions of Grx2 outside mitochondria.