Intrinsically Disordered Regions Promote Protein Refoldability and Facilitate Retrieval from Biomolecular Condensates

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: June 25, 2023

Abstract Many eukaryotic proteins contain intrinsically disordered regions (IDRs) that intersperse globular folded domains, in contrast with bacterial which are typically highly 1, 2 . Recent years have seen great progress identifying biological functions associated these elusive protein sequence: specific cases, they mediate liquid phase separation 3 , perform molecular recognition 4 or act as sensors to changes the environment 5 Nevertheless, only a small number of IDRs annotated 6 despite their presence 64% yeast proteins, 7 stimulating some question what ‘general purpose’ may serve 8, 9 Here, by interrogating refoldability two fungal proteomes ( Saccharomyces cerevisiae and Neurosporra crassa ), we show render host more refoldable from denatured state, allowing them cohere closely Anfinsen’s thermodynamic hypothesis 10, 11 The data provide an exceptionally clear picture biophysical topological characteristics enable refoldability. Moreover, find almost all partition into stress granules during heat shock refoldable, finding holds for other condensates such P-bodies nucleolus. Finally, Hsp104 unfoldase 12 is principal actor mediating disassembly efficiency condensed returned soluble also well explained Hence, studies establish spontaneous adaptive trait endows capacity reform native structures following extraction condensates. Altogether, our results intuitive model function many multidomain clarifies relationship phenomenon biomolecular condensation.

Language: Английский

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Protein aggregation and biomolecular condensation in hypoxic environments (Review)

International Journal of Molecular Medicine, Journal Year: 2024, Volume and Issue: 53(4)

Published: Feb. 12, 2024

Due to molecular forces, biomacromolecules assemble into liquid condensates or solid aggregates, and their corresponding formation dissolution processes are controlled. Protein homeostasis is disrupted by increasing age environmental stress, leading irreversible protein aggregation. Hypoxic pressure an important factor in this process, uncontrolled aggregation has been widely observed hypoxia‑related conditions such as neurodegenerative disease, cardiovascular hypoxic brain injury cancer. Biomolecular also high‑order complexes assembled from macromolecules. Although they exist different phase dynamic balance under certain conditions, activation assembly considered regulatory cell survival with pressure. Therefore, a better understanding of the relationship between biomolecular condensation will bring marked benefits clinical treatment various diseases. The aim present review was summarize underlying mechanisms aggregate induced address recent breakthroughs role aggregates hypoxic‑related diseases, given hypotheses that hypoxia induces macromolecular assemblage changes phase, adenosine triphosphate depletion ATP‑driven inactivation multiple chaperones play roles among process. Moreover, it anticipated improved adaptation environments could extend overall patients provide new strategies for

Language: Английский

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Isoform-specific sequestration of protein kinase A fine-tunes intracellular signaling during heat stress

Cell Reports, Journal Year: 2024, Volume and Issue: 43(6), P. 114360 - 114360

Published: June 1, 2024

Protein kinase A (PKA) is a conserved crucial for fundamental biological processes linked to growth, development, and metabolism. The PKA catalytic subunit expressed as multiple isoforms in diverse eukaryotes; however, their contribution ensuring signaling specificity response environmental cues remains poorly defined. Catalytic activity classically moderated via interaction with an inhibitory regulatory subunit. Here, quantitative mass spectrometry approach used examine heat-stress-induced changes the binding of yeast Tpk1–3 subunits Bcy1 We show that Tpk3 not regulated by but, instead, deactivated upon heat stress reversible sequestration into cytoplasmic granules. These "Tpk3 granules" are enriched substrates involved various metabolic processes, Hsp42 sequestrase required formation. Hence, provides mechanism control isoform-specific during conditions.

Language: Английский

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Melatonin: Regulation of Viral Phase Separation and Epitranscriptomics in Post-Acute Sequelae of COVID-19

International Journal of Molecular Sciences, Journal Year: 2022, Volume and Issue: 23(15), P. 8122 - 8122

Published: July 23, 2022

The relentless, protracted evolution of the SARS-CoV-2 virus imposes tremendous pressure on herd immunity and demands versatile adaptations by human host genome to counter transcriptomic epitranscriptomic alterations associated with a wide range short- long-term manifestations during acute infection post-acute recovery, respectively. To promote viral replication active persistence, envelope protein regulates cell microenvironment including pH ion concentrations maintain high oxidative environment that supports template switching, causing extensive mitochondrial damage activation pro-inflammatory cytokine signaling cascades. Oxidative stress distress induce dynamic changes both RNA m

Language: Английский

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Artificial Hsp104-mediated systems for re-localizing protein aggregates

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: May 9, 2023

Abstract Spatial Protein Quality Control (sPQC) sequesters misfolded proteins into specific, organelle-associated inclusions within the cell to control their toxicity. To approach role of sPQC in cellular fitness, neurodegenerative diseases and aging, we report on construction Hsp100-based systems budding yeast cells, which can artificially target protein aggregates non-canonical locations. We demonstrate that mutant huntingtin (mHtt), disease-causing agent Huntington’s disease be targeted daughter cells as well eisosomes endosomes with this approach. find artificial removal mHtt from mother protects them death suggesting even large may cytotoxic, a trait has been widely debated. In contrast, removing endogenous age-associated does not significantly affect lifespan cells. also is able manipulate inclusion formation human potential useful an alternative, complementary study sPQC, for example aging disease.

Language: Английский

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Genetic inactivation of essential HSF1 reveals an isolated transcriptional stress response selectively induced by protein misfolding

Molecular Biology of the Cell, Journal Year: 2023, Volume and Issue: 34(10)

Published: July 19, 2023

Heat Shock Factor 1 (Hsf1) in yeast drives the basal transcription of key proteostasis factors and its activity is induced as part core heat shock response. Exploring Hsf1 specific functions has been challenging due to essential nature HSF1 gene extensive overlap target promoters with environmental stress response (ESR) Msn2 Msn4 (Msn2/4). In this study, we constructed a viable hsf1∆ strain by replacing open reading frame genes that constitutively express Hsp40, Hsp70, Hsp90 from Hsf1-independent promoters. Phenotypic analysis showed grows slowly, sensitive well protein misfolding accumulates aggregates. Transcriptome revealed transcriptional azetidine-2-carboxylic acid fully dependent on Hsf1. contrast, responded through ESR. Following HS, Msn2/4 functional compensatory induction stronger activation remaining pathway when other branch was inactivated. Thus, provide long-overdue genetic test function using novel construct. Our data highlight accumulation misfolded proteins uniquely sensed Hsf1-Hsp70 chaperone titration inducing highly selective

Language: Английский

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mRNA condensation fluidizes the cytoplasm

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: May 31, 2023

The intracellular environment is packed with macromolecules of mesoscale size, and this crowded milieu significantly influences cell physiology. When exposed to stress, mRNAs released after translational arrest condense RNA binding proteins, resulting in the formation membraneless protein (RNP) condensates known as processing bodies (P-bodies) stress granules (SGs). However, impact assembly these on biophysical properties cytoplasmic remains unclear. Here, we find that upon exposure polysome collapse condensation increases particle diffusivity cytoplasm. Increased required for efficient Q-bodies, organelles coordinate degradation misfolded peptides accumulate during stress. Additionally, demonstrate granule has a similar effect mammalian cells, fluidizing cytoplasm at mesoscale. We synthetic, light-induced sufficient fluidize cytoplasm, demonstrating causal condensation. Together, our work reveals new functional role stress-induced translation inhibition RNP modulating physical effectively respond stressful conditions.

Language: Английский

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Protein aggregation: A detrimental symptom or an adaptation mechanism?

Journal of Neurochemistry, Journal Year: 2023, Volume and Issue: 168(8), P. 1426 - 1441

Published: Sept. 11, 2023

Abstract Protein quality control mechanisms oversee numerous aspects of protein lifetime. From the point synthesis, homeostasis machineries take part in folding, solubilization, and/or degradation impaired proteins. Some proteins follow an alternative path upon loss their solubility, thus are secluded from cytosol and form aggregates. aggregates differ function composition, rendering aggregation a complex phenomenon that continues to receive plenty attention scientific medical communities. Traditionally, have been associated with aging large spectrum folding diseases, such as neurodegenerative type 2 diabetes, or cataract. However, body evidence suggests they may act adaptive mechanism overcome transient stressful conditions, serving sink for removal misfolded storage compartments required stress release. In this review, we present examples elaborating different possible roles discuss potential survival, aging, disease, well anti‐aggregation interventions. image

Language: Английский

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Spatial sequestration of misfolded proteins in neurodegenerative diseases

Biochemical Society Transactions, Journal Year: 2022, Volume and Issue: 50(2), P. 759 - 771

Published: March 21, 2022

Properly folded, functional proteins are essential for cell health. Cells sustain protein homeostasis, or proteostasis, via quality control (PQC) mechanisms. It is currently hypothesized that a breakdown in proteostasis during ageing leads to the accumulation of aggregates and disease. Sequestration misfolded into PQC compartments represents one branch network. In neurodegenerative diseases, certain form abnormal deposits. Which house associated with diseases still being investigated. remains unclear if sequestration these toxic protective cell. Here, we review current knowledge on various cell, kinds found what known about their sequestration. Understanding how occurs can shed light why linked like Huntington's, Alzheimer's, Parkinson's diseases.

Language: Английский

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Solid-to-liquid phase transition in the dissolution of cytosolic misfolded-protein aggregates

iScience, Journal Year: 2023, Volume and Issue: 26(12), P. 108334 - 108334

Published: Oct. 28, 2023

Accumulation of protein aggregates is a hallmark cellular aging and degenerative disorders. This could result from either increased misfolding aggregation or impaired dissolution formed under stress, the latter which poorly understood. In this study, we employed quantitative live-cell imaging to investigate dynamic process disaggregation in yeast. We show that upon heat stress are solid condensates, but after attenuation these first transition into liquid-like state during their dissolution. solid-to-liquid phase (SLPT) accompanies reduction aggregate number due fusion liquid condensates. The chaperone activity Hsp104, Clp/HSP100 family chaperone, required for both SLPT subsequent dispersal Sse1, yeast HSP110 also facilitates SLPT. These results illuminate an unexpected mechanistic framework control over attenuation.

Language: Английский

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