Hsp70 and Hsp90 of E. coli Directly Interact for Collaboration in Protein Remodeling

Genest, Olivier; Hoskins, Joel R.; Kravats, Andrea N.; Doyle, Shannon M.; Wickner, Sue

doi:10.1016/j.jmb.2015.10.010

Cited by 70 publications

(123 citation statements)

References 55 publications

Supporting

Mentioning

105

Contrasting

Order By: Relevance

“…In addition to ATP, TRAP1-dependent protein folding also requires the functional cooperation of the mitochondrial Hsp70 system. The latter supports a direct physical interaction, as was recently reported for cytosolic Hsp90 chaperones (5)(6)(7)50), and opens up new avenues for drug development by targeting the specific interaction between mitochondrial chaperones.…”

Section: Discussionsupporting

confidence: 83%

Mitochondrial Hsp90 is a ligand-activated molecular chaperone coupling ATP binding to dimer closure through a coiled-coil intermediate

Sung

Lee

Kim

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Heat-shock protein of 90 kDa (Hsp90) is an essential molecular chaperone that adopts different 3D structures associated with distinct nucleotide states: a wide-open, V-shaped dimer in the apo state and a twisted, N-terminally closed dimer with ATP. Although the N domain is known to mediate ATP binding, how Hsp90 senses the bound nucleotide and facilitates dimer closure remains unclear. Here we present atomic structures of human mitochondrial Hsp90 N (TRAP1 N ) and a composite model of intact TRAP1 revealing a previously unobserved coiled-coil dimer conformation that may precede dimer closure and is conserved in intact TRAP1 in solution. Our structure suggests that TRAP1 normally exists in an autoinhibited state with the ATP lid bound to the nucleotide-binding pocket. ATP binding displaces the ATP lid that signals the cis-bound ATP status to the neighboring subunit in a highly cooperative manner compatible with the coiled-coil intermediate state. We propose that TRAP1 is a ligand-activated molecular chaperone, which couples ATP binding to dramatic changes in local structure required for protein folding.TRAP1 | Hsp90 | molecular chaperone H eat-shock protein of 90 kDa (Hsp90) is a conserved ATPdependent molecular chaperone (1-4), which together with heat-shock protein of 70 kDa (Hsp70) (5-7) and a cohort of cochaperones (8-10), promotes the late-stage folding of Hsp90 client proteins (11). It is presumed that almost 400 different proteins, including a majority of signaling and tumor promoting proteins, depend on cytosolic Hsp90 for folding (12). Consequently, the ability to inactivate multiple oncogenic pathways simultaneously has made Hsp90 a major target for drug development (13), with several Hsp90 inhibitors currently undergoing clinical trials (14).Hsp90 chaperones display conformational plasticity in solution (2, 15, 16), with different adenine nucleotides either facilitating or stabilizing distinct Hsp90 dimer conformations (17-19). Interestingly, apo Hsp90 forms a wide-open, V-shaped dimer with the N domains separated by as much as 101 Å (18). This open conformation is markedly distinct from the intertwined, N-terminally closed dimer with ATP bound (20,21). Because the open-state dimer cannot signal the nucleotide status between neighboring subunits, an intermediate conformation preceding dimer closure must exist, which so far has remained elusive.Apart from cytosolic Hsp90s, Hsp90 homologs are found in the endoplasmic reticulum, chloroplasts, and mitochondria ( Fig. S1) (22). The tumor necrosis factor receptor-associated protein 1 (TRAP1) is the mitochondrial Hsp90 paralog, which prevents apoptosis and protects mitochondria against oxidative damage (23-25). TRAP1 is widely expressed in many tumors (24,26,27), but not in mitochondria of most normal tissues (24), benign prostatic hyperplasia (26), or highly proliferating, nontransformed cells (27). Notably, it was found that TRAP1 not only promotes neoplastic growth, but also confers tumorigenic potential on nontransformed cells (27), indicating a major rol...

show abstract

Section: Discussionsupporting

confidence: 83%

Mitochondrial Hsp90 is a ligand-activated molecular chaperone coupling ATP binding to dimer closure through a coiled-coil intermediate

Sung

Lee

Kim

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…We previously showed that E. coli DnaK collaborates with Hsp90 Ec in protein reactivation and that the two chaperones physically interact through a region in the middle domain of Hsp90 Ec [23, 30]. To elucidate the region of DnaK essential for binding Hsp90 Ec , we used molecular docking to predict potential interactions between the two chaperones.…”

Section: Resultsmentioning

confidence: 99%

“…GrpE, the bacterial NEF stimulates reactivation, but is not essential [23]. ATP hydrolysis and client binding by both chaperones is essential for reactivation [30]. Moreover, bacterial Hsp90 physically interacts with DnaK in vivo and in vitro [29, 30] through a region identified on the M-domain of Hsp90 Ec [30].…”

Section: Introductionmentioning

confidence: 99%

Interaction of E. coli Hsp90 with DnaK Involves the DnaJ Binding Region of DnaK

Kravats

Doyle

Hoskins

et al. 2017

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

Hsp90 is a widely conserved and ubiquitous molecular chaperone that participates in ATP-dependent protein remodeling in both eukaryotes and prokaryotes. It functions in conjunction with Hsp70 and the Hsp70 cochaperones, an Hsp40 (J-protein) and a nucleotide exchange factor. In E. coli the functional collaboration between Hsp90Ec and Hsp70, DnaK, requires that the two chaperones directly interact. We used molecular docking to model the interaction of Hsp90Ec and DnaK. The top-ranked docked model predicted that a region in the nucleotide-binding domain of DnaK interacted with a region in the middle domain of Hsp90Ec. We then made substitution mutants in DnaK residues suggested by the model to interact with Hsp90Ec. Eleven of the twelve mutants tested were defective or partially defective in their ability to interact with Hsp90Ec in vivo in a bacterial two-hybrid assay and in vitro in a Bio-Layer Interferometry assay. These DnaK mutants were also defective in their ability to function collaboratively in protein remodeling with Hsp90Ec, but retained the ability to act with DnaK cochaperones. Taken together these results suggest that a specific region in the nucleotide-binding domain of DnaK is involved in the interaction with Hsp90Ec and this interaction is functionally important. Moreover, the region of DnaK that we found to be necessary for Hsp90Ec binding includes residues that are also involved in J-protein binding, suggesting a functional interplay between DnaK, DnaK cochaperones and Hsp90Ec.

show abstract

“…Within the chaperone network, many of the major chaperones make physical interactions with each other. For example, the prokaryotic orthologs of Hsp70 and Hsp90 directly bind to each other [9; 10], as do the orthologs of Hsp70 and Hsp110 [11]. In mammals, Hsp70s are physically linked to the Hsp90s, but through the scaffolding protein, HOP (Hsc70-Hsp90 organizing factor).…”

Section: Introductionmentioning

confidence: 99%

BAG3 Is a Modular, Scaffolding Protein that physically Links Heat Shock Protein 70 (Hsp70) to the Small Heat Shock Proteins

Rauch¹,

Tse

Freilich³

et al. 2017

Journal of Molecular Biology

118

133

View full text Add to dashboard Cite

Small heat shock proteins (sHsps) are a family of ATP-independent molecular chaperones that are important for binding and stabilizing unfolded proteins. In this task, the sHsps have been proposed to coordinate with ATP-dependent chaperones, including heat shock protein 70 (Hsp70). However, it isn’t yet clear how these two important components of the chaperone network are linked. We report that the Hsp70 co-chaperone, BAG3, is a modular, scaffolding factor to bring together sHsps and Hsp70s. Using domain deletions and point mutations, we confirmed that BAG3 uses both of its IPV motifs to interact with sHsps, including Hsp27 (HspB1), αB-crystallin (HspB5), Hsp22 (HspB8) and Hsp20 (HspB6). BAG3 does not appear to be a passive scaffolding factor; rather, its binding promoted de-oligomerization of Hsp27, likely by competing for the self-interactions that normally stabilize large oligomers. BAG3 bound to Hsp70 at the same time as either Hsp22, Hsp27 or αB-crystallin, suggesting that it might physically bring the chaperone families together into a complex. Indeed, addition of BAG3 coordinated the ability of Hsp22 and Hsp70 to refold denatured luciferase in vitro. Together, these results suggest that BAG3 physically and functionally links Hsp70 and sHsps.

show abstract

Hsp70 and Hsp90 of E. coli Directly Interact for Collaboration in Protein Remodeling

Cited by 70 publications

References 55 publications

Mitochondrial Hsp90 is a ligand-activated molecular chaperone coupling ATP binding to dimer closure through a coiled-coil intermediate

Mitochondrial Hsp90 is a ligand-activated molecular chaperone coupling ATP binding to dimer closure through a coiled-coil intermediate

Interaction of E. coli Hsp90 with DnaK Involves the DnaJ Binding Region of DnaK

BAG3 Is a Modular, Scaffolding Protein that physically Links Heat Shock Protein 70 (Hsp70) to the Small Heat Shock Proteins

Contact Info

Product

Resources

About