Interactions of the chaperone Hsp104 with yeast Sup35 and mammalian PrP

Schirmer, Eric C.; Lindquist, Susan

doi:10.1073/pnas.94.25.13932

Cited by 79 publications

(56 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…5,35 However, one study by Schirmer and Lindquist showed a weak inhibition by Aβ 42 with a reduction by a factor of 0.8. 23 This discrepancy might be due to differences in the aggregation state of Aβ or assay conditions such as salt content, pH, the presence of traces of denaturants (guanidinium chloride), and peptide concentration, to which Hsp104 is highly sensitive. 6,34 Beyond the ATPase assays, which are an indirect measure of the substrate interaction, direct methods were employed to study the binding of Aβ to Hsp104.…”

Section: Binding Of Aβ Can Be Monitored By Atp Turnover Of Hsp104 Andmentioning

confidence: 99%

Hsp104 Targets Multiple Intermediates on the Amyloid Pathway and Suppresses the Seeding Capacity of Aβ Fibrils and Protofibrils

Arimon

Grimminger

Sanz

et al. 2008

Journal of Molecular Biology

View full text Add to dashboard Cite

The heat shock protein Hsp104 has been reported to possess the ability to modulate protein aggregation and toxicity and to "catalyze" the disaggregation and recovery of protein aggregates, including amyloid fibrils, in yeast, Escherichia coli, mammalian cell cultures, and animal models of Huntington's disease and Parkinson's disease. To provide mechanistic insight into the molecular mechanisms by which Hsp104 modulates aggregation and fibrillogenesis, the effect of Hsp104 on the fibrillogenesis of amyloid beta (Aβ) was investigated by characterizing its ability to interfere with oligomerization and fibrillogenesis of different species along the amyloid-formation pathway of Aβ. To probe the disaggregation activity of Hsp104, its ability to dissociate preformed protofibrillar and fibrillar aggregates of Aβ was assessed in the presence and in the absence of ATP. Our results show that Hsp104 inhibits the fibrillization of monomeric and protofibrillar forms of Aβ in a concentration-dependent but ATP-independent manner. Inhibition of Aβ fibrillization by Hsp104 is observable up to Hsp104/Aβ stoichiometric ratios of 1:1000, suggesting a preferential interaction of Hsp104 with aggregation intermediates (e.g., oligomers, protofibrils, small fibrils) on the pathway of Aβ amyloid formation. This hypothesis is consistent with our observations that Hsp104 (i) interacts with Aβ protofibrils, (ii) inhibits conversion of protofibrils into amyloid fibrils, (iii) arrests fibril elongation and reassembly, and (iv) abolishes the capacity of protofibrils and sonicated fibrils to seed the fibrillization of monomeric Aβ. Together, these findings suggest that the strong inhibition of Aβ fibrillization by Hsp104 is mediated by its ability to act at different stages and target multiple intermediates on the pathway to amyloid formation.

show abstract

Section: Binding Of Aβ Can Be Monitored By Atp Turnover Of Hsp104 Andmentioning

confidence: 99%

Hsp104 Targets Multiple Intermediates on the Amyloid Pathway and Suppresses the Seeding Capacity of Aβ Fibrils and Protofibrils

Arimon

Grimminger

Sanz

et al. 2008

Journal of Molecular Biology

View full text Add to dashboard Cite

show abstract

“…The C-terminal region of Sup35p contains the essential translation activity (Ter-Avanesyan et al, 1993), and the N-terminal region confers upon Sup35p the capacity to assume distinct prion and nonprion conformations. The change between these states is regulated by interactions between the N-terminal domain and Hsp104p (Patino et al, 1996;Schirmer and Lindquist, 1997;Cashikar et al, 2002). When Sup35p prion conversion is too efficient it can be toxic (Ter-Avanesyan et al, 1993;Derkatch et al, 1996;Li and Lindquist, 2000), because the essential translation-termination activity of the C-terminal domain is inhibited when the protein is in the [PSI ϩ ] state.…”

Section: Molecular Biology Of the Cell 2064mentioning

confidence: 99%

Dominant Gain-of-Function Mutations in Hsp104p Reveal Crucial Roles for the Middle Region

Schirmer

Homann

Kowal

et al. 2004

MBoC

Self Cite

106

111

View full text Add to dashboard Cite

Heat-shock protein 104 (Hsp104p) is a protein-remodeling factor that promotes survival after extreme stress by disassembling aggregated proteins and can either promote or prevent the propagation of prions (protein-based genetic elements). Hsp104p can be greatly overexpressed without slowing growth, suggesting tight control of its powerful protein-remodeling activities. We isolated point mutations in Hsp104p that interfere with this control and block cell growth. Each mutant contained alterations in the middle region (MR). Each of the three MR point mutations analyzed in detail had distinct phenotypes. In combination with nucleotide binding site mutations, Hsp104p T499I altered bud morphology and caused septin mislocalization, colocalizing with the misplaced septins. Point mutations in the septin Cdc12p suppressed this phenotype, suggesting that it is due to direct Hsp104p-septin interactions. Hsp104p A503V did not perturb morphology but stopped cell growth. Remarkably, when expressed transiently, the mutant protein promoted survival after extreme stress as effectively as did wild-type Hsp104p. Hsp104p A509D had no deleterious effects on growth or morphology but had a greatly reduced ability to promote thermotolerance. That mutations in an 11-amino acid stretch of the MR have such profound and diverse effects suggests the MR plays a central role in regulating Hsp104p function. INTRODUCTIONThe AAA ϩ proteins are ATPases associated with various cellular activities. They are important proteins with a great diversity of functions, including protein folding, membrane trafficking, organelle biogenesis, proteolysis, intracellular motility, and DNA replication (Neuwald et al., 1999;Vale, 2000;Ogura and Wilkinson, 2001). Little is known about how most AAA ϩ proteins recognize substrates and use ATP binding and hydrolysis to remodel them. The intrinsic complexity and multiplicity of conformational states of the AAA ϩ proteins make them difficult to study. The Clp/ HSP100 proteins are members of the AAA ϩ superfamily and have been the subject of intense biochemical analysis in vitro and genetic analysis in vivo (Wickner et al., 1999;Glover and Tkach, 2001). The functional unit of the yeast HSP100 heatshock protein 104 (Hsp104p) is composed of six monomers, each with two ATP binding sites (nucleotide binding domains one and two; NBD1 and NBD2) flanked by aminoterminal, middle, and carboxy-terminal regions (Figure 1).Hsp104p has remarkable functions, one of which is to allow survival after extreme stress. For example, yeast cells expressing Hsp104p are 1000 times more viable after exposure to temperatures Ն50°C or to an ethanol concentration of 20% than cells carrying deletions of HSP104 (Sanchez and Lindquist, 1990;Sanchez et al., 1992). This survival capacity is directly attributable to Hsp104p's ability to resolubilize protein aggregates and, together with Hsp70p and Hsp40p, return them to their folded and active states (Parsell et al., 1994;Glover and Lindquist, 1998;Goloubinoff et al., 1999). This is in contrast to other...

show abstract

“…Importantly for the current study, genetic experiments have clearly shown that the HSPs regulate assembly of both prions and polyQ fragments (2, 3, 5, 19 -21). For example, Hsp70 activity appears to be required to facilitate Hsp104 disaggregase activity during the propagation and inheritance of prions (22)(23)(24) and other model substrates (25). In addition, Hsp26 works with Hsp104 to help break large fibrils into inheritable fragments (26), while Hsp70 and Hsp90 also assist in this process (27).…”

mentioning

confidence: 99%

Ordered Assembly of Heat Shock Proteins, Hsp26, Hsp70, Hsp90, and Hsp104, on Expanded Polyglutamine Fragments Revealed by Chemical Probes

Walter

Smith

Wisén

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

In Saccharomyces cerevisae, expanded polyglutamine (polyQ) fragments are assembled into discrete cytosolic aggregates in a process regulated by the molecular chaperones Hsp26, Hsp70, Hsp90, and Hsp104. To better understand how the different chaperones might cooperate during polyQ aggregation, we used sequential immunoprecipitations and mass spectrometry to identify proteins associated with either soluble (Q25) or aggregation-prone (Q103) fragments at both early and later times after induction of their expression. We found that Hsp26, Hsp70, Hsp90, and other chaperones interact with Q103, but not Q25, within the first 2 h. Further, Hsp70 and Hsp90 appear to be partially released from Q103 prior to the maturation of the aggregates and before the recruitment of Hsp104. To test the importance of this seemingly ordered process, we used a chemical probe to artificially enhance Hsp70 binding to Q103. This treatment retained both Hsp70 and Hsp90 on the polyQ fragment and, interestingly, limited subsequent exchange for Hsp26 and Hsp104, resulting in incomplete aggregation. Together, these results suggest that partial release of Hsp70 may be an essential step in the continued processing of expanded polyQ fragments in yeast.The heat shock proteins (HSPs) 3 are abundant molecular chaperones that have been shown to be important for maintaining proteostasis under both normal conditions and during times of cellular stress (1-3). Together, these factors play multiple roles in quality control, especially related to polypeptide synthesis, folding, and turnover (4 -7). Moreover, HSPs are emerging as drug targets in cancer, neurodegenerative disorders and other diseases (8, 9). Thus, there is interest in better understanding how they coordinate protein quality control decisions.The major families of HSPs are named based on their approximate kDa molecular weights, including Hsp60, Hsp70, Hsp90,

show abstract

Interactions of the chaperone Hsp104 with yeast Sup35 and mammalian PrP

Cited by 79 publications

References 32 publications

Hsp104 Targets Multiple Intermediates on the Amyloid Pathway and Suppresses the Seeding Capacity of Aβ Fibrils and Protofibrils

Hsp104 Targets Multiple Intermediates on the Amyloid Pathway and Suppresses the Seeding Capacity of Aβ Fibrils and Protofibrils

Dominant Gain-of-Function Mutations in Hsp104p Reveal Crucial Roles for the Middle Region

Ordered Assembly of Heat Shock Proteins, Hsp26, Hsp70, Hsp90, and Hsp104, on Expanded Polyglutamine Fragments Revealed by Chemical Probes

Contact Info

Product

Resources

About