Structural visualization of the tubulin folding pathway directed by human chaperonin TRiC/CCT

Gestaut, Daniel R.; Zhao, Yanyan; Junsun, Park; Ma, Boxue; Leitner, Alexander; Collier, M; Pintilie, G.; Roh, Sook Young; Chiu, Wah; Frydman, Judith

doi:10.1016/j.cell.2022.11.014

Cited by 37 publications

(28 citation statements)

References 79 publications

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“…We also find that the PhLP2A CTD is extended from the TXD which proximally locates to the substrate binding site at the inter-chamber space of TRiC. The TXD crosslinks to the flexible CCT tails, which contribute to substrate binding 10 , 28 , 41 . Therefore, it is tempting to speculate that the PhLP2A TXD may sense the substrate directly via the proximal interaction and cascades the binding affinity to the CTD.…”

Section: Discussionmentioning

confidence: 75%

“…A recent structure of TRiC directly captured from cell lysate showed that it could form a ternary complex with actin and PhLP2A 28 . We added PhLP2A to a reconstituted TRiC-mediated actin folding reaction using purified components and a physiologically relevant system 10 . Briefly, human PFD-bound actin remains in a high protease K sensitive unstructured state (Figure S6A).…”

Section: Resultsmentioning

confidence: 99%

“…Upon lid closure, the encapsulated substrate interacts extensively with the closed wall as well as the N- and C-tails. In this context, the fact that PhLP2A domains bind to multiple sites inside the chamber in both open and closed TRiC states is remarkable, particularly in light of recent findings showing interactions with the TRiC chamber direct the folding trajectories of at least some substrates such as tubulin and actin 4 , 10 . Our findings suggest that PhLPs must modulate the interactions and/or conformation of the TRiC-bound substrate inside the chamber, and possibly the TRiC conformational cycle itself.…”

Section: Discussionmentioning

confidence: 99%

“…TRiC facilitates the folding of ~10% of the proteome and diversifies the strategies to support folding for many different types of protein substrates. The heterooligomeric nature of TRiC is considered key to diversifying the recognition of distinct motifs in its diverse substrates 4 , 5 , 46 , as well as promoting their folding within the closed chamber 4 , 10 . Our analysis of PhLP family evolution suggests the modality of binding of family members to the TRiC chamber is largely conserved, whereas the substrate binding elements within the NTD, and perhaps the CTD, have diversified to assist folding to different substrate proteins.…”

Section: Discussionmentioning

confidence: 99%

“…Prefoldin is a jellyfish-shaped hetero-oligomeric complex that binds the open state of TRiC through specific contacts with the apical domains but is released upon ATP-driven lid closure 15 . PFD maintains substrates in a dynamic unstructured conformation 10 and enhances the processivity of TRiC-mediated folding by transferring substrates to the open TRiC cavity 15 . The PhLPs are a family of ~30 kDa cytosolic proteins that have also been shown to regulate TRiC-mediated protein folding 16 .…”

Section: Introductionmentioning

confidence: 99%

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A structural vista of phosducin-like PhLP2A-chaperonin TRiC cooperation during the ATP-driven folding cycle

Junsun

Kim

Gestaut

et al. 2023

Preprint

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Proper cellular proteostasis, essential for viability, requires a network of chaperones and cochaperones. ATP-dependent chaperonin TRiC/CCT partners with cochaperones prefoldin (PFD) and phosducin-like proteins (PhLPs) to facilitate the folding of essential eukaryotic proteins. Using cryoEM and biochemical analyses, we determine the ATP-driven cycle of TRiC-PFD-PhLP2A interaction. In the open TRiC state, PhLP2A binds to the chamber's equator while its N-terminal H3-domain binds to the apical domains of CCT3/4, thereby displacing PFD from TRiC. ATP-induced TRiC closure rearranges the contacts of PhLP2A domains within the closed chamber. In the presence of substrate, actin and PhLP2A segregate into opposing chambers, each binding to the positively charged inner surfaces formed by CCT1/3/6/8. Notably, actin induces a conformational change in PhLP2A, causing its N-terminal helices to extend across the inter-ring interface to directly contact a hydrophobic groove in actin. Our findings reveal an ATP-driven PhLP2A structural rearrangement cycle within the TRiC chamber to facilitate folding.

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Section: Discussionmentioning

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A structural vista of phosducin-like PhLP2A-chaperonin TRiC cooperation during the ATP-driven folding cycle

Junsun

Kim

Gestaut

et al. 2023

Preprint

Self Cite

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Revisiting the chaperonin T‐complex protein‐1 ring complex in human health and disease: A proteostasis modulator and beyond

Zeng,

Han,

Pan

et al. 2024

Clinical & Translational Med

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BackgroundDisrupted protein homeostasis (proteostasis) has been demonstrated to facilitate the progression of various diseases. The cytosolic T‐complex protein‐1 ring complex (TRiC/CCT) was discovered to be a critical player in orchestrating proteostasis by folding eukaryotic proteins, guiding intracellular localisation and suppressing protein aggregation. Intensive investigations of TRiC/CCT in different fields have improved the understanding of its role and molecular mechanism in multiple physiological and pathological processes.Main bodyIn this review, we embark on a journey through the dynamic protein folding cycle of TRiC/CCT, unraveling the intricate mechanisms of its substrate selection, recognition, and intriguing folding and assembly processes. In addition to discussing the critical role of TRiC/CCT in maintaining proteostasis, we detail its involvement in cell cycle regulation, apoptosis, autophagy, metabolic control, adaptive immunity and signal transduction processes. Furthermore, we meticulously catalogue a compendium of TRiC‐associated diseases, such as neuropathies, cardiovascular diseases and various malignancies. Specifically, we report the roles and molecular mechanisms of TRiC/CCT in regulating cancer formation and progression. Finally, we discuss unresolved issues in TRiC/CCT research, highlighting the efforts required for translation to clinical applications, such as diagnosis and treatment.ConclusionThis review aims to provide a comprehensive view of TRiC/CCT for researchers to inspire further investigations and explorations of potential translational possibilities.

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In silico protein dynamics in the human cytoplasm: Partial folding, misfolding, fold switching, and non‐native interactions

Russell,

Rickard,

Boob

et al. 2023

Protein Science

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We examine the influence of cellular interactions in all‐atom models of a section of the Homo sapiens cytoplasm on the early folding events of the three‐helix bundle protein B (PB). While genetically engineered PB is known to fold in dilute water box simulations in three microseconds, the three initially unfolded PB copies in our two cytoplasm models using a similar force field did not reach the native state during 30 microsecond simulations. We did however capture formation of all three helices in a compact native‐like topology. Folding in vivo is delayed because intramolecular contact formation within PB is in direct competition with intermolecular contacts between PB and surrounding macromolecules. In extreme cases, intermolecular beta‐sheets are formed. Interactions with other macromolecules is also observed to promote structure formation, for example when a PB helix in our simulations is shielded from solvent by macromolecular crowding. Sticking and crowding in our models initiate sampling of helix/sheet structural plasticity of PB. Relatedly, in past in vitro experiments, similar GA domains were shown to switch between two different folds. Finally, we also observed that stickiness between PB and the cellular environment can be modulated in our simulations through reduction in protein hydrophobicity when we reversed PB back to the wild type sequence. This study demonstrates that even fast‐folding proteins can get stuck in non‐native states in the cell, making them useful models for protein‐chaperone interactions and early stages of aggregate formation relevant to cellular disease.This article is protected by copyright. All rights reserved.

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Structural visualization of the tubulin folding pathway directed by human chaperonin TRiC/CCT

Cited by 37 publications

References 79 publications

A structural vista of phosducin-like PhLP2A-chaperonin TRiC cooperation during the ATP-driven folding cycle

A structural vista of phosducin-like PhLP2A-chaperonin TRiC cooperation during the ATP-driven folding cycle

Revisiting the chaperonin T‐complex protein‐1 ring complex in human health and disease: A proteostasis modulator and beyond

In silico protein dynamics in the human cytoplasm: Partial folding, misfolding, fold switching, and non‐native interactions

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