Potentiated Hsp104 variants antagonize diverse proteotoxic misfolding events.
There are no therapies that reverse the proteotoxic misfolding events that underpin fatal neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD). Hsp104, a conserved hexameric AAA+ protein from yeast, solubilizes disordered aggregates and amyloid but has no metazoan homolog and only limited activity against human neurodegenerative disease proteins. Here, we reprogram Hsp104 to rescue TDP-43, FUS, and α-synuclein proteotoxicity by mutating single residues in helix 1, 2, or 3 of the middle domain or the small domain of nucleotide-binding domain 1. Potentiated Hsp104 variants enhance aggregate dissolution, restore proper protein localization, suppress proteotoxicity, and in a C. elegans PD model attenuate dopaminergic neurodegeneration. Potentiating mutations reconfigure how Hsp104 subunits collaborate, desensitize Hsp104 to inhibition, obviate any requirement for Hsp70, and enhance ATPase, translocation, and unfoldase activity. Our work establishes that disease-associated aggregates and amyloid are tractable targets and that enhanced disaggregases can restore proteostasis and mitigate neurodegeneration.
Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Meredith E
ME
Jackrel
Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA; Biochemistry and Molecular Biophysics Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Morgan E
ME
DeSantis
Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
Bryan A
BA
Martinez
Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA; Pharmacology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Laura M
LM
Castellano
Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Rachel M
RM
Stewart
Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
Kim A
KA
Caldwell
Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
Guy A
GA
Caldwell
Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA; Biochemistry and Molecular Biophysics Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA; Pharmacology Graduate Group, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: jshorter@mail.med.upenn.edu.
James
J
Shorter
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D004268:DNA-Binding Proteins / Q000378:metabolism; D004195:Disease Models, Animal; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D009474:Neurons / Q000166:cytology / Q000473:pathology; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D057165:Proteostasis Deficiencies / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D034702:RNA-Binding Protein FUS / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051844:alpha-Synuclein / Q000378:metabolism
D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D034702:RNA-Binding Protein FUS; D029701:Saccharomyces cerevisiae Proteins; D051844:alpha-Synuclein; C070972:HsP104 protein, S cerevisiae