title: Vulnerability of newly synthesized proteins to proteostasis stress.
abstract: 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 ∼5-10% of the soluble cytosolic and nuclear proteome in human HEK293 cells is vulnerable to misfolding when proteostatic function is overwhelmed. Inhibiting new protein synthesis for 30 min prior to heat-shock dramatically reduced the amount of heat-stress induced polyubiquitylation, and reduced the misfolding of proteins identified as vulnerable to thermal stress. Following prior studies in C. elegans in 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.
authors.affiliation: Department of Neuroscience, SantaFe HealthCare Alzheimer's Disease Research Center, Center for Translational Research in Neurodegenerative Disease, McKnight Brain Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Guilian
authors.initials: G
authors.lastname: Xu
authors.affiliation: Interdisciplinary Graduate Program, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Amrutha
authors.initials: A
authors.lastname: Pattamatta
authors.affiliation: College of Arts and Sciences, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Hildago
authors.affiliation: Interdisciplinary Graduate Program, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Pace
authors.affiliation: Department of Neuroscience, SantaFe HealthCare Alzheimer's Disease Research Center, Center for Translational Research in Neurodegenerative Disease, McKnight Brain Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Hilda
authors.initials: H
authors.lastname: Brown
authors.affiliation: Department of Neuroscience, SantaFe HealthCare Alzheimer's Disease Research Center, Center for Translational Research in Neurodegenerative Disease, McKnight Brain Institute, University of Florida, Gainesville, FL 32610, USA drb1@ufl.edu.
authors.forename: David R
authors.initials: DR
authors.lastname: Borchelt
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D000071058:Huntingtin Protein / Q000235:genetics / Q000378:metabolism*; D020125:Mutation, Missense; D066263:Protein Aggregation, Pathological / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014176:Protein Biosynthesis; D057165:Proteostasis Deficiencies / Q000235:genetics / Q000378:metabolism* / Q000473:pathology
chemical_list: D029742:Caenorhabditis elegans Proteins; D000071058:Huntingtin Protein
keywords: Heat-shock; Neurodegenerative disease; Protein aggregation; Proteomics; Proteostasis; Ubiquitin