Mechanistic and Structural Insights into the Prion-Disaggregase Activity of Hsp104

Sweeny, Elizabeth A.; Shorter, James

doi:10.1016/j.jmb.2015.11.016

Cited by 85 publications

(115 citation statements)

References 169 publications

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“…By applying different heterohexamer ensembles composed of Hsp104 and Hsp104 A503V subunits to reactivate disordered luciferase aggregates, we can obtain a measure of how many A503V subunits per hexamer are required for disaggregase activity in the absence of Hsp70 (Fig. 1, B and C) (26,29,61,62,70).…”

Section: Hsp104 Hexamers Must Contain 2-3 A503v Subunits Formentioning

confidence: 99%

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Mechanistic Insights into Hsp104 Potentiation

Torrente¹,

Chuang

Noll³

et al. 2016

Journal of Biological Chemistry

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Potentiated variants of Hsp104, a protein disaggregase from yeast, can dissolve protein aggregates connected to neurodegenerative diseases such as Parkinson disease and amyotrophic lateral sclerosis. However, the mechanisms underlying Hsp104 potentiation remain incompletely defined. Here, we establish that 2-3 subunits of the Hsp104 hexamer must bear an A503V potentiating mutation to elicit enhanced disaggregase activity in the absence of Hsp70. We also define the ATPase and substratebinding modalities needed for potentiated Hsp104 A503V activity in vitro and in vivo. Hsp104 A503V disaggregase activity is strongly inhibited by the Y257A mutation that disrupts substrate binding to the nucleotide-binding domain 1 (NBD1) pore loop and is abolished by the Y662A mutation that disrupts substrate binding to the NBD2 pore loop. A503V displays a more robust activity that is unperturbed by sensor-1 mutations that greatly reduce Hsp104 activity in vivo. Indeed, ATPase activity at NBD1 or NBD2 is sufficient for Hsp104 potentiation. Our findings will empower design of ameliorated therapeutic disaggregases for various neurodegenerative diseases.Protein misfolding and aggregation are associated with a wide variety of diseases, ranging from type II diabetes (1, 2) to neurodegenerative diseases, such as fatal familial insomnia (3, 4), Parkinson disease, and amyotrophic lateral sclerosis (ALS) (5-7). In Parkinson disease patients, ␣-synuclein (␣-syn) 6 forms toxic soluble oligomers as well as amyloid structures that accumulate in Lewy bodies and contribute to the death of dopaminergic neurons (8 -12). Similarly, toxic soluble oligomers and cytoplasmic inclusions of TDP-43 or FUS are associated with ALS and frontotemporal dementia (13-20). These misfolded protein conformers are recalcitrant and represent a colossal roadblock in the treatment of these diseases.Hsp104 is a 102-kDa AAAϩ ATPase (21) from Saccharomyces cerevisiae capable of dissolving disordered protein aggregates as well as dismantling amyloid fibrils and toxic soluble oligomers (22-33). It assembles into a homohexameric barrel structure with a central channel (34 -39). Hsp104 processes protein aggregates by directly translocating substrates either partially or completely through this channel (35, 36, 40 -46). Hsp104 encompasses an N-terminal domain, two nucleotidebinding domains (NBD1 and NBD2), a coiled-coil middle domain (MD), and a C-terminal domain important for oligomerization (Fig. 1A) (47). Both NBDs contain Walker A and Walker B motifs that are critical for nucleotide binding and hydrolysis, respectively (48). ATP hydrolysis takes place primarily at NBD1, whereas NBD2 has a nucleotide-dependent oligomerization function (29, 34, 49 -52).Remarkably, Hsp104 can remodel amyloid substrates alone, without the aid of any other chaperones (22,24,26,31,33,45,(53)(54)(55)(56). However, to disaggregate amorphous protein aggregates, Hsp104 usually needs to collaborate with the Hsp110, Hsp70, and Hsp40 chaperone system (23,26,30,32,57). Moreover, small heat shock proteins...

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Section: Hsp104 Hexamers Must Contain 2-3 A503v Subunits Formentioning

confidence: 99%

“…Hsp104 is a 102-kDa AAAϩ ATPase (21) from Saccharomyces cerevisiae capable of dissolving disordered protein aggregates as well as dismantling amyloid fibrils and toxic soluble oligomers (22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33). It assembles into a homohexameric barrel structure with a central channel (34 -39).…”

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confidence: 99%

Mechanistic Insights into Hsp104 Potentiation

Torrente¹,

Chuang

Noll³

et al. 2016

Journal of Biological Chemistry

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“…What makes one-dimensional polymers able to act as prions is that they, unlike liquids, crystals, or glasses, can be fragmented by point applications of force. Even a single protein -- most prominently the AAA+ ATPase Hsp104 -- pulling on a single monomer within the amyloid suffices to snap it into two pieces [72]. Most characterized prions require Hsp104 activity; in its absence, the amyloids fail to fragment and hence cannot transmit to other cells.…”

Section: Figurementioning

confidence: 99%

A glass menagerie of low complexity sequences

Halfmann

2016

Current Opinion in Structural Biology

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Remarkably simple proteins play outsize roles in the execution of developmental complexity within biological systems. Sequence information determines structure and hence function, so how do low complexity sequences fulfill their functions? Recent discoveries are raising the curtain on a new dimension of the sequence-structure paradigm. In it, function derives not from the structures of individual proteins, but instead, from dynamic material properties of entire ensembles of the proteins acting in unison through phase changes. These phases include liquids, one-dimensional crystals, and -- as elaborated herein -- even glasses. The peculiar thermodynamics of glass-like protein assemblies, in particular, illuminate new principles of information flow through and, at times, orthogonal to the central dogma of molecular biology.

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“…Hsp104 is an asymmetric ring-shaped translocase and hexameric AAA+ protein found in yeast [30,31••]. Hsp104 couples ATP hydrolysis to the rapid dissolution and reactivation of diverse proteins trapped in disordered aggregates, ordered stress-induced assemblies, preamyloid oligomers, amyloids, and prions [32–37,38••].…”

Section: Hsp104 a Protein Disaggregase From Yeastmentioning

confidence: 99%

Designer protein disaggregases to counter neurodegenerative disease

Shorter

2017

Current Opinion in Genetics & Development

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Protein misfolding and aggregation unify several devastating neurodegenerative disorders, including Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis. There are no effective therapeutics for these disorders and none that target the reversal of the aberrant protein misfolding and aggregation that cause disease. Here, I showcase important advances to define, engineer, and apply protein disaggregases to mitigate deleterious protein misfolding and counter neurodegeneration. I focus on two exogenous protein disaggregases, Hsp104 from yeast and gene 3 protein from bacteriophages, as well as endogenous human protein disaggregases, including: (a) Hsp110, Hsp70, Hsp40, and small heat-shock proteins; (b) HtrA1; and (c) NMNAT2 and Hsp90. I suggest that protein-disaggregase modalities can be channeled to treat numerous fatal and presently incurable neurodegenerative diseases.

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Mechanistic and Structural Insights into the Prion-Disaggregase Activity of Hsp104

Cited by 85 publications

References 169 publications

Mechanistic Insights into Hsp104 Potentiation

Mechanistic Insights into Hsp104 Potentiation

A glass menagerie of low complexity sequences

Designer protein disaggregases to counter neurodegenerative disease

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