Mining Disaggregase Sequence Space to Safely Counter TDP-43, FUS, and α-Synuclein Proteotoxicity

Tariq, Amber; Lin, Jia Bei; Jackrel, Meredith E.; Hesketh, Christina D.; Carman, Peter J.; Mack, Korrie L.; Weitzman, Rachel; Gambogi, Craig W.; Murillo, Oscar Hernandez; Sweeny, Elizabeth A.; Gurpinar, Esin; Yokom, Adam L.; Gates, S.N.; Yee, Keolamau; Sudesh, Saurabh; Stillman, Jacob M.; Rizo, Alexandra N.; Southworth, Daniel R.; Shorter, James

doi:10.1016/j.celrep.2019.07.069

Cited by 36 publications

(94 citation statements)

References 88 publications

(180 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3b,c, S6b,c,d). These results demonstrate that Skd3 disaggregase activity is regulated by PARL and that PARL-activated Skd3 is a powerful, stand-alone protein disaggregase with comparable activity to potentiated Hsp104 variants Jackrel et al, 2015;Tariq et al, 2019;Tariq et al, 2018).…”

Section: Skd3 Disaggregase Activity Is Enhanced By Parl Cleavagementioning

confidence: 66%

Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations

Cupo

Shorter

2020

Preprint

Self Cite

View full text Add to dashboard Cite

Cells have evolved specialized protein disaggregases to reverse toxic protein aggregation and restore protein functionality. In nonmetazoan eukaryotes, the AAA+ disaggregase Hsp78 resolubilizes and reactivates proteins in mitochondria. Curiously, metazoa lack Hsp78. Hence, whether metazoan mitochondria reactivate aggregated proteins is unknown. Here, we establish that a mitochondrial AAA+ protein, Skd3 (human CLPB), couples ATP hydrolysis to protein disaggregation and reactivation. The Skd3 ankyrin-repeat domain combines with conserved AAA+ elements to enable stand-alone disaggregase activity. A mitochondrial inner-membrane protease, PARL, removes an autoinhibitory peptide from Skd3 to greatly enhance disaggregase activity. Indeed, PARL-activated Skd3 dissolves α-synuclein fibrils connected to Parkinson's disease. Human cells lacking Skd3 exhibit reduced solubility of various mitochondrial proteins, including anti-apoptotic Hax1. Importantly, Skd3 variants linked to 3-methylglutaconic aciduria, a severe mitochondrial disorder, display diminished disaggregase activity (but not always reduced ATPase activity), which predicts disease severity. Thus, Skd3 is a potent protein disaggregase critical for human health.

show abstract

Section: Skd3 Disaggregase Activity Is Enhanced By Parl Cleavagementioning

confidence: 66%

Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations

Cupo

Shorter

2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, we did not isolate any Hsp104 homologs capable of suppressing FUS toxicity. In contrast, potentiated Hsp104 variants typically are able to suppress toxicity of multiple toxic substrates (Figures 1, 2, and S3) [30,[32][33][34]37]. Natural Hsp104 homologs also displayed no intrinsic toxicity, even when expressed at elevated temperatures ( Figure S2).…”

Section: Discussionmentioning

confidence: 99%

“…Thus, extant Hsp104 homologs have likely been filtered through natural selection to avoid deleterious and destabilizing sequences. In contrast, potentiated Hsp104 variants have predominantly been engineered by destabilizing the NBD1:MD interface [30,[33][34][35]37]. This difference is further reflected in fundamental mechanistic differences between how the Hsp104 homologs described here and potentiated Hsp104 variants operate to antagonize proteotoxic misfolding.…”

Section: Discussionmentioning

confidence: 99%

“…In yeast, galactose-inducible expression of several proteins associated with neurodegenerative diseases, including aSyn [39], TDP-43 [40], and FUS [13] is toxic. However, coexpression of potentiated Hsp104 variants or the natural homolog Calcarisporiella thermophila Hsp104 (CtHsp104), but not ScHsp104, reduces this toxicity [27,30,33,37]. For a deeper exploration of natural Hsp104 Table S1 for homolog sequences, Figures S1 for an alignment of all homologs).…”

Section: Diverse Hsp104 Homologs Selectively Suppress Tdp-43 Toxicitymentioning

confidence: 99%

“…However, in some circumstances, these Hsp104 variants can also exhibit off-target toxicity [30,32,33]. Thus, uncovering other therapeutic Hsp104s with diminished propensity for off-target effects is a key objective [37].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Therapeutic genetic variation revealed in diverse Hsp104 homologs

March

Sweeney

Kim

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

The AAA+ protein disaggregase, Hsp104, increases fitness under stress by reversing stress-induced protein aggregation. We have engineered potentiated Hsp104 variants to antagonize proteotoxic misfolding linked to human neurodegenerative diseases. However, these Hsp104 variants can exhibit offtarget toxicity, which may limit their therapeutic utility. Hsp104 is conserved among all nonmetazoan eukaryotes, which raises the possibility that natural variants might exist with enhanced, selective activity against neurodegenerative disease substrates. To assess this possibility, we screened a cross-kingdom collection of Hsp104 homologs in several yeast proteotoxicity models. We uncovered therapeutic genetic variation among several Hsp104 homologs that specifically antagonize TDP-43 or a-synuclein condensate formation and toxicity in yeast, human cells, and C. elegans. Surprisingly, this variation manifested as increased passive chaperone activity, distinct from disaggregase activity, which neutralizes proteotoxicity of specific substrates. Thus, by exploring natural tuning of passive chaperone activity we elucidated enhanced, substrate-specific agents to counter proteotoxicity underlying neurodegenerative disease.

show abstract

The clinical trial landscape in amyotrophic lateral sclerosis—Past, present, and future

Wobst

Mack

Brown

et al. 2020

Medicinal Research Reviews

View full text Add to dashboard Cite

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease marked by progressive loss of muscle function. It is the most common adult‐onset form of motor neuron disease, affecting about 16 000 people in the United States alone. The average survival is about 3 years. Only two interventional drugs, the antiglutamatergic small‐molecule riluzole and the more recent antioxidant edaravone, have been approved for the treatment of ALS to date. Therapeutic strategies under investigation in clinical trials cover a range of different modalities and targets, and more than 70 different drugs have been tested in the clinic to date. Here, we summarize and classify interventional therapeutic strategies based on their molecular targets and phenotypic effects. We also discuss possible reasons for the failure of clinical trials in ALS and highlight emerging preclinical strategies that could provide a breakthrough in the battle against this relentless disease.

show abstract

Mining Disaggregase Sequence Space to Safely Counter TDP-43, FUS, and α-Synuclein Proteotoxicity

Cited by 36 publications

References 88 publications

Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations

Skd3 (human CLPB) is a potent mitochondrial protein disaggregase that is inactivated by 3-methylglutaconic aciduria-linked mutations

Therapeutic genetic variation revealed in diverse Hsp104 homologs

The clinical trial landscape in amyotrophic lateral sclerosis—Past, present, and future

Contact Info

Product

Resources

About