The capacity of the cell to produce, fold, and degrade proteins relies on components of the proteostasis network. Multiple types of insults can impose a burden on this network, causing protein misfolding. Using thermal stress, a classic example of acute proteostatic stress, we demonstrate that about 5-10% of the soluble cytosolic/nuclear proteome in human 293 cells is vulnerable to misfolding when proteostatic function is over-whelmed. Inhibiting new protein synthesis for 30 minutes prior to heat-shock dramatically reduced the amount of heat-stress induced polyubiquitination, and reduced the misfolding of proteins identified as vulnerable to thermal stress. Following prior studies inC. elegansin which mutant huntingtin (Q103) expression was shown to cause the secondary misfolding of cytosolic proteins, we also demonstrate that mutant huntingtin causes similar “secondary” misfolding in human cells. Similar to thermal stress, inhibiting new protein synthesis reduced the impact of mutant huntingtin on proteostatic function. These findings suggest that newly made proteins are vulnerable to misfolding when proteostasis is disrupted by insults such as thermal stress and mutant protein aggregation.
© 2016. Published by The Company of Biologists Ltd.