Quantifying Nucleation In Vivo Reveals the Physical Basis of Prion-like Phase Behavior

Khan, Tarique; Kandola, Tejbir S.; Wu, Jianzheng; Somasundaram, Venkatesan; Ketter, Ellen; Lange, Jeffrey J.; Gama, Alejandro Rodriguez; Box, Andrew; Unruh, Jay R.; Cook, Malcolm; Halfmann, Randal

doi:10.1016/j.molcel.2019.01.045

Cited by 8 publications

(2 citation statements)

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“…We produced cell lines expressing WT tau fused to CFP which were treated with CBD patient material to replicate the CBD conformation in the cells. Subsequently, we transduce tau fused to mEOS3.2 72 encoding alanine mutations to test the ability of each mutation to incorporate into the WT-propagated CBD-derived inclusions in the cells. Mutations that interfere with incorporation into CBD-derived inclusions should not yield FRET between CFP and mEOS3.2 while mutations that can incorporate into inclusions should have similar FRET to WT tau.…”

Section: Identification Of Thermodynamic Hotspots Of Aggregationmentioning

confidence: 99%

Network of hotspot interactions cluster tau amyloid folds

et al. 2023

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Cryogenic electron microscopy has revealed unprecedented molecular insight into the conformations of β-sheet-rich protein amyloids linked to neurodegenerative diseases. It remains unknown how a protein can adopt a diversity of folds and form multiple distinct fibrillar structures. Here we develop an in silico alanine scan method to estimate the relative energetic contribution of each amino acid in an amyloid assembly. We apply our method to twenty-seven ex vivo and in vitro fibril structural polymorphs of the microtubule-associated protein tau. We uncover networks of energetically important interactions involving amyloid-forming motifs that stabilize the different fibril folds. We evaluate our predictions in cellular and in vitro aggregation assays. Using a machine learning approach, we classify the structures based on residue energetics to identify distinguishing and unifying features. Our energetic profiling suggests that minimal sequence elements control the stability of tau fibrils, allowing future design of protein sequences that fold into unique structures.

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Section: Identification Of Thermodynamic Hotspots Of Aggregationmentioning

confidence: 99%

Network of hotspot interactions cluster tau amyloid folds

et al. 2023

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“…It has been suggested that the heterotypic interactions between [ PIN + ] and the high concentration of Sup35NM through overexpression fosters amyloid nucleation and bypasses the formation of condensates. When [ PIN + ] is absent, it is well established that prion formation is diminished and the overexpression of Sup35NM leads to condensate formation rather than the prion state (Khan et al, ).…”

Section: Sup35p As a Prionmentioning

confidence: 99%

The three faces of Sup35

Lyke

Dorweiler

Manogaran

2019

Yeast

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Sup35p is an essential protein in yeast that functions in complex with Sup45p for efficient translation termination. Although some may argue that this function is the only important attribute of Sup35p, there are two additional known facets of Sup35p's biology that may provide equally important functions for yeast; both of which involve various strategies for coping with stress. Recently, the N‐terminal and middle regions (NM) of Sup35p, which are not required for translation termination function, have been found to provide stress‐sensing abilities and facilitate the phase separation of Sup35p into biomolecular condensates in response to transient stress. Interestingly, the same NM domain is also required for Sup35p to misfold and enter into aggregates associated with the [PSI+] prion. Here, we review these three different states or “faces” of Sup35p. For each, we compare the functionality and necessity of different Sup35p domains, including the role these domains play in facilitating interactions with important protein partners, and discuss the potential ramifications that each state affords yeast cells under varying environmental conditions.

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