Structure of the RAF1-HSP90-CDC37 complex reveals the basis of RAF1 regulation

García-Alonso, Sara; Mesa, Pablo; Ovejero, Laura de la Puente; Aizpurua, Gonzalo; Lechuga, Carmen G.; Zarzuela, Eduardo; Santiveri, Clara M.; Sanclemente, Manuel; Muñoz, Javier; Musteanu, Mónica; Campos-Olivas, Ramón; Martı́nez-Torrecuadrada, Jorge L.; Barbacid, Mariano; Montoya, Guillermo

doi:10.1016/j.molcel.2022.08.012

Cited by 26 publications

(27 citation statements)

References 59 publications

(65 reference statements)

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“…In this map, Hsp90 sidechains could be readily interpreted and more detail for Cdc37 and the kinase was observed than in our previous, lower-resolution Hsp90:Cdc37:Cdk4 kinase complex structure 33 . As expected from the Cdk4 kinase complex, and further seen in the recent CRaf kinase complex 42 , the CRaf KD kinase domain in our structure was split into its distinct lobes, threaded through the Hsp90 lumen and stabilized by Cdc37. This new map shows a closed Hsp90, with the CRaf KD C-lobe extending from the Hsp90 lumen on one side while the prominent Cdc37 N-terminal coiled-coil projects away from the opposite complex surface.…”

Section: Resultssupporting

confidence: 82%

Hsp90 provides a platform for kinase dephosphorylation by PP5

et al. 2023

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The Hsp90 molecular chaperone collaborates with the phosphorylated Cdc37 cochaperone for the folding and activation of its many client kinases. As with many kinases, the Hsp90 client kinase CRaf is activated by phosphorylation at specific regulatory sites. The cochaperone phosphatase PP5 dephosphorylates CRaf and Cdc37 in an Hsp90-dependent manner. Although dephosphorylating Cdc37 has been proposed as a mechanism for releasing Hsp90-bound kinases, here we show that Hsp90 bound kinases sterically inhibit Cdc37 dephosphorylation indicating kinase release must occur before Cdc37 dephosphorylation. Our cryo-EM structure of PP5 in complex with Hsp90:Cdc37:CRaf reveals how Hsp90 both activates PP5 and scaffolds its association with the bound CRaf to dephosphorylate phosphorylation sites neighboring the kinase domain. Thus, we directly show how Hsp90’s role in maintaining protein homeostasis goes beyond folding and activation to include post translationally modifying its client kinases.

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Section: Resultssupporting

confidence: 82%

Hsp90 provides a platform for kinase dephosphorylation by PP5

et al. 2023

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“…The Cricetulus griseus HSP90β and Cdc37 show remarkable sequence conservation in comparison to the human equivalents, at 99.7 and 94.2% identity, respectively. This native pulldown strategy contrasts with the structures of Hsp90–Cdc37 in complex with soluble kinases (García-Alonso et al, 2022; Oberoi et al, 2022; Verba et al, 2016), for which Hsp90 and Cdc37 had to be overexpressed to obtain complex suitable for imaging. Three-dimensional reconstruction of our GC-C–Hsp90–Cdc37 particles generated a 3.9 Å resolution map of the regulatory complex (Figure 1, Supplementary Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…The present cryoEM structure of GC-C–Hsp90–Cdc37 resolves the loading of GC-C, via its PK domain and interaction with Cdc37, to the Hsp90 core dimer (Figure 1, 2). This complex shows significant structural similarity to the mechanism used to regulate soluble active kinases (García-Alonso et al, 2022; Oberoi et al, 2022; Verba et al, 2016) and presumably membrane receptor kinases in the human proteome. This structural and mechanistic conservation is largely driven by the co-chaperone Cdc37, which serves as the main binding platform for these clients by associating to the fold of the kinase (or pseudokinase in the case of mGC) domain, relatively independent of sequence identity.…”

Section: Discussionmentioning

confidence: 97%

“…The core structural principles of Cdc37 mediated client recruitment to Hsp90 appear to remain constant across its large range of client diversity. Across other client-Hsp90-Cdc37 complexes with canonical soluble kinase clients (Cdk4, RAF1, B-raf) (García-Alonso et al, 2022;Oberoi et al, 2022;Verba et al, 2016), we see a conserved role for Cdc37 in client recruitment by associating with the C-lobe at the N-, C-lobe interface (Supplemental Figure 2A). In these complexes we see high levels of structural conservation for the Hsp90-Cdc37 (Cα RMSDs of 1.4-3.3 Å for Hsp90 and 1.5-2.5 Å for Cdc37), while the client is structurally most homogenous at the interface with Cdc37, though less structurally conserved overall (Cα RMSDs of 3.5-11.6 Å).…”

Section: Conservation Of Cdc37 Mediated Hsp90 Regulationmentioning

confidence: 90%

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Structural insight into guanylyl cyclase receptor hijacking of the kinase–Hsp90 regulatory mechanism

Caveney

Tsutsumi

Garcia

2023

Preprint

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Membrane receptor guanylyl cyclases play a role in many important facets of human physiology, ranging from regulation of blood pressure to the regulation of intestinal fluid secretion. The structural mechanisms which influence the regulation of these important physiological effects have yet to be explored. We present the 3.9 A resolution cryoEM structure of the human membrane receptor guanylyl cyclase GC-C in complex with Hsp90 and its co-chaperone Cdc37, providing insight into the mechanism of Cdc37 mediated binding of GC-C to the Hsp90 regulatory complex. As a membrane protein and non-kinase client of Hsp90-Cdc37, this work shows remarkable plasticity of Cdc37 to interact with a broad array of clients having significant sequence variation. Further, this work shows how membrane receptor guanylyl cyclases hijack the regulatory mechanisms used for active kinases to facilitate their regulation. Given the known druggability of Hsp90, these insights can guide the further development of mGC targeted therapeutics and lead to new avenues to treat hypertension, inflammatory bowel disease, and other mGC related conditions.

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“…BRAF is a cytosolic serine/threonine kinase belonging to the RAF kinase family, serving as an important step of signal transmission from the cell surface to the nucleus after EGFR activation [122]. As a part of MAPK pathway, BRAF is involved in cell growth, proliferation, survival, and differentiation [123]. BRAF mutations occur in 1.5%-3.5% of NSCLC cases.…”

Section: Braf Mutationsmentioning

confidence: 99%

Advances in Targeted Therapy Against Driver Mutations and Epigenetic Alterations in Non-Small Cell Lung Cancer

Shi¹,

Chen²,

Peng³

et al. 2022

Oncologie

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The incidence and mortality of lung cancer rank top three of all cancers worldwide. Accounting for 85% of the total number of lung cancer, non-small cell lung cancer (NSCLC) is an important factor endangering human health. Recently, targeted therapies against driver mutations and epigenetic alterations have made encouraging advances that benefit NSCLC patients. Druggable driver mutations, which mainly occur in EGFR, KRAS, MET, HER2, ALK, ROS1, RET and BRAF, have been identified in more than a quarter of NSCLC patients. A series of highly selective mutant targeting inhibitors, such as EGFR tyrosine kinase inhibitors and KRAS inhibitors, have been well studied and applied in clinical treatments, which greatly promote the overall survival of NSCLC patients. However, drug resistance has become a major challenge for targeted treatment, and a variety of methods to overcome drug resistance are constantly being developed, including inhibitors against new mutants, combination therapy with other pathway inhibitors, etc. In addition, epigenetics-based therapy is emerging. Epigenetic regulators such as histone deacetylases and non-coding RNA play a crucial role in the development of cancer and drug resistance by affecting multiple signaling pathways. Epigenetics-based therapeutic strategies combined with targeted drugs show great clinical potential. Many agents targeting epigenetic changes are being investigated in preclinical studies, with some already under clinical trials. This article focuses on driver mutations and epigenetic alterations in association with relevant epidemiological data. We introduce the current status of targeted inhibitors and known drug resistance, review advances in major targeted therapies with recent data from preclinical and clinical trials, and discuss the possibility of combination therapy against driver mutations and epigenetic alterations in overcoming drug resistance.

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Structure of the RAF1-HSP90-CDC37 complex reveals the basis of RAF1 regulation

Cited by 26 publications

References 59 publications

Hsp90 provides a platform for kinase dephosphorylation by PP5

Hsp90 provides a platform for kinase dephosphorylation by PP5

Structural insight into guanylyl cyclase receptor hijacking of the kinase–Hsp90 regulatory mechanism

Advances in Targeted Therapy Against Driver Mutations and Epigenetic Alterations in Non-Small Cell Lung Cancer

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