The Hsp40 J‐domain modulates Hsp70 conformation and ATPase activity with a semi‐elliptical spring

Bascos, Neil Andrew D.; Mayer, Mathias; Bukau, Bernd; Landry, Samuel J.

doi:10.1002/pro.3223

Cited by 20 publications

(19 citation statements)

References 43 publications

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“…HSP40 belongs to DNAJ family subcategorized into three subclasses, which are DnaJA (DNAJA), DnaJB (DNAJB), and DnaJC (DNAJC) [6]. HSP40 assists in protein folding, unfolding, translation, translocation, and degradation [11,25,26], as well as ATPase activity of HSP70 [27]. Many of the HSP40 family members are overexpressed in numerous human cancer types, such as colorectal, gastric, and lung cancers [28][29][30][31].…”

Section: Oncogenic Role Of Hsp40 In Proliferation and Metastasis Of Cmentioning

confidence: 99%

Heat Shock Proteins: Agents of Cancer Development and Therapeutic Targets in Anti-Cancer Therapy

Yun

Kim

Lim

et al. 2019

Cells

210

158

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Heat shock proteins (HSPs) constitute a large family of molecular chaperones classified by their molecular weights, and they include HSP27, HSP40, HSP60, HSP70, and HSP90. HSPs function in diverse physiological and protective processes to assist in maintaining cellular homeostasis. In particular, HSPs participate in protein folding and maturation processes under diverse stressors such as heat shock, hypoxia, and degradation. Notably, HSPs also play essential roles across cancers as they are implicated in a variety of cancer-related activities such as cell proliferation, metastasis, and anti-cancer drug resistance. In this review, we comprehensively discuss the functions of HSPs in association with cancer initiation, progression, and metastasis and anti-cancer therapy resistance. Moreover, the potential utilization of HSPs to enhance the effects of chemo-, radio-, and immunotherapy is explored. Taken together, HSPs have multiple clinical usages as biomarkers for cancer diagnosis and prognosis as well as the potential therapeutic targets for anti-cancer treatment.

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Section: Oncogenic Role Of Hsp40 In Proliferation and Metastasis Of Cmentioning

confidence: 99%

Heat Shock Proteins: Agents of Cancer Development and Therapeutic Targets in Anti-Cancer Therapy

Yun

Kim

Lim

et al. 2019

Cells

210

158

View full text Add to dashboard Cite

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“…However, we do not mean to say that this role of the D HPD is the only mechanistic effect of J-domain binding to Hsp70. For example, the invariant histidine and/or proline of the HPD may play specific roles, and, as previously suggested 39 , conformational strain induced in the J-domain upon binding to Hsp70 might be important as well. However, there is no question that R NBD is key and could be considered as an allosteric perturbation site for the Jdomain action.…”

Section: Discussionmentioning

confidence: 75%

Two-step mechanism of J-domain action in driving Hsp70 function

Tomiczek

Delewski

Nierzwicki

et al. 2020

Preprint

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J-domain proteins (JDPs), obligatory Hsp70 cochaperones, play critical roles in protein homeostasis. They promote key allosteric transitions that stabilize Hsp70 interaction with substrate polypeptides upon hydrolysis of its bound ATP. Although a recent crystal structure revealed the physical mode of interaction between a J-domain and an Hsp70, the structural and dynamic consequences of J-domain action once bound and how Hsp70s discriminate among its multiple JDP partners remain enigmatic. We combined free energy simulations, biochemical assays and evolutionary analyses to address these issues. Our results indicate that the invariant aspartate of the J-domain perturbs a conserved intramolecular Hsp70 network of contacts that crosses domains. This perturbation leads to destabilization of the domain-domain interface -thereby promoting the allosteric transition that triggers ATP hydrolysis. While this mechanistic step is driven by conserved residues, evolutionarily variable residues are key to initial JDP/Hsp70 recognition -via electrostatic interactions between oppositely charged surfaces. We speculate that these variable residues allow an Hsp70 to discriminate amongst JDP partners, as many of them have coevolved. Together, our data points to a two-step mode of J-domain action, a recognition stage followed by a mechanistic stage. are able to discriminate amongst their multiple JDP partners, thus enabling evolution of complex JDP/Hsp70 interaction networks. ResultsStructural model of the Hsc20-Ssq1 complex. To obtain a structural model of the JDP-Hsp70 complex formed by Hsc20 and Ssq1, we combined protein-protein docking with all-atom molecular dynamics (MD) simulations ( Supplementary Fig. S1). This approach revealed a dominant bound state that accounted for 56% of the bound population ( Fig. 1, Supplementary Fig. S2a). No other bound state accounted for more than 6% of the ensemble; these minor states rapidly interconverted and had a clear tendency to converge to the most populated bound state ( Supplementary Fig. S2b). The binding mode in this dominant state closely resembles the recently published X-ray structure of DnaJ's J-domain in complex with DnaK (DnaJ JD -DnaK) 22 . Helices II and III are oriented very similarly in the two complexes -with a backbone root mean square deviation (RMSD) of 4.17 Å ( Supplementary Fig. S3). More specifically, the J-domains bind at the NBD/SBDb,linker interface such that helix II predominantly interacts with the b-sheet region of NBD subdomain IIa and helix III with SBDb (Fig. 1). The HPD residues are positioned nearly identically in the two complexes, towards R167/R207 of NBD subdomain Ia in DnaK/Ssq1 (referred to as R NBD throughout), the arginine that previous analyses of DnaK established as important for allosteric transitions 14,20,21 . D HPD interferes in Hsp70 interdomain interactions by perturbing critical R NBD contacts.To address the question of the mechanism of J-domain action, we employed MD simulations that, unlike most other methods, allow studying of transient r...

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“…Despite their structural diversity, they all share a well-conserved J domain (Jd), which has been found to be crucial for their interaction with Hsp70s (5). Altered interactions between the Hsp40 Jd and Hsp70s, both in terms of binding affinity (6,7), and mechanical properties (8), have been related to changes in the ability of Jd to promote Hsp70 functions. A recent report on the structure of a Jd docked to an Hsp70 molecule in E. coli revealed a network of polar and nonpolar interactions proposed to transmit the signal from the Jd to the catalytic center at the NBD (9).…”

Section: Introductionmentioning

confidence: 99%

Structural influence of the conserved Hsp40 HPD tripeptide on Hsp70 chaperone function

Mallapre

Bascos

2020

Preprint

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The control of Hsp70 functions has been related to the modulation of ATP hydrolysis and substrate capture by Hsp40. Structural and biophysical analyses of Hsp40 variants and their interactions with Hsp70 have identified key residues for this functional control mechanism. Conserved residues in both Hsp40 and Hsp70 have revealed conserved interactions that link Hsp40 binding to the catalytic residues within Hsp70. The current work investigates the effect of documented J-domain dysfunctional mutations (i.e. D35N, H33Q) on the described interaction linkage. Molecular dynamics simulations were used to compare the persistence of individual bond types (i.e. H-bonds, salt bridges, hydrophobic interactions) between Hsp70 and the bound forms of functional and dysfunctional Hsp40 variants. The generated data suggests the involvement of both direct and allosteric effects for the tested mutations. The observed changes relate mutations in the conserved HPD tripeptide of Hsp40 to alterations in the interaction network that induces Hsp70 chaperone functions.The significance of the work may be summarized as follows. First, the interaction network for the simulated systems were observed to be different from one previously proposed for a disulfide linked complex (9). This may be attributed to altered residue movement and interactions without the restrictions set by the disulfide link. These results support the use of in silico methods to refine investigations of molecular contacts, particularly for systems, whose in vitro structural elucidation are difficult to achieve without modifications.Second, key interactions for intermolecular and intramolecular contacts were observed within a short simulation time (0.1 ns) matched those from much longer runs (500 ns) (4). This result highlights the possibility of identifying key interactions with relatively low computational cost.

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The Hsp40 J‐domain modulates Hsp70 conformation and ATPase activity with a semi‐elliptical spring

Cited by 20 publications

References 43 publications

Heat Shock Proteins: Agents of Cancer Development and Therapeutic Targets in Anti-Cancer Therapy

Heat Shock Proteins: Agents of Cancer Development and Therapeutic Targets in Anti-Cancer Therapy

Two-step mechanism of J-domain action in driving Hsp70 function

Structural influence of the conserved Hsp40 HPD tripeptide on Hsp70 chaperone function

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