Balanced activities of Hsp70 and the ubiquitin proteasome system underlie cellular protein homeostasis DOI Creative Commons
Areeb Jawed,

Chi‐Ting Ho,

Tomáš Groušl

et al.

Frontiers in Molecular Biosciences, Journal Year: 2023, Volume and Issue: 9

Published: Jan. 4, 2023

To counteract proteotoxic stress and cellular aging, protein quality control (PQC) systems rely on the refolding, degradation sequestration of misfolded proteins. In Saccharomyces cerevisiae Hsp70 chaperone system plays a central role in while is predominantly executed by ubiquitin proteasome (UPS). The sequestrases Hsp42 Btn2 deposit proteins cytosolic nuclear inclusions, thereby restricting accessibility to preventing exhaustion limited resources. Therefore, yeast, sequestrase mutants show negative genetic interactions with double lacking co-chaperone Fes1 Hsp104 disaggregase ( fes1Δ hsp104Δ , ΔΔ ) suffering from low capacity. Growth ΔΔbtn2Δ highly temperature-sensitive results proteostasis breakdown at non-permissive temperatures. Here, we probed for maintaining homeostasis cells, which are affected two major branches. We that cells induce expression diverse stress-related pathways including defects. Ubiquitin dependent stringent substrate firefly Luciferase mutant mirrors such compensatory activities system. Surprisingly however, enhanced activity does not improve but aggravates growth defects cells. Reducing lowering levels functional 26S proteasomes improved growth, increased refolding yield reporter attenuated global responses. Our findings indicate an imbalance between Hsp70-dependent system-mediated strongly affects capacity contributes their severe phenotypes.

Language: Английский

Cellular Control of Viscosity Counters Changes in Temperature and Energy Availability DOI Creative Commons

Laura Persson,

Vardhaan S. Ambati, Onn Brandman

et al.

Cell, Journal Year: 2020, Volume and Issue: 183(6), P. 1572 - 1585.e16

Published: Nov. 5, 2020

Language: Английский

Citations

149
Mechanisms tailoring the expression of heat shock proteins to proteostasis challenges DOI Creative Commons
Lokha R. Alagar Boopathy, Suleima Jacob‐Tomas, Célia Alecki

et al.

Journal of Biological Chemistry, Journal Year: 2022, Volume and Issue: 298(5), P. 101796 - 101796

Published: March 3, 2022

All cells possess an internal stress response to cope with environmental and pathophysiological challenges. Upon stress, reprogram their molecular functions activate a survival mechanism known as the heat shock response, which mediates rapid induction of chaperones such proteins (HSPs). This potent production overcomes general suppression gene expression results in high levels HSPs subsequently refold or degrade misfolded proteins. Once damage is repaired removed, terminate resume regular functions. Thus, fulfillment requires swift robust coordination between activation completion that determined by status cell. In recent years, single-cell fluorescence microscopy techniques have begun be used unravelling HSP-gene pathways, from DNA transcription mRNA degradation. this review, we will address mechanisms different organisms cell types coordinate signaling networks act transcription, translation, decay ensure protein quality control. For grow function properly, they must maintain specific cellular conditions allow acquire functional conformations achieve homeostasis (proteostasis) (1Gasch A.P. Spellman P.T. Kao C.M. Carmel-Harel O. Eisen M.B. Storz G. Botstein D. Brown P.O. Genomic programs yeast changes.Mol. Biol. Cell. 2000; 11: 4241-4257Crossref PubMed Google Scholar). Maintaining proteostasis becomes critical when facing abrupt changes external conditions, increase temperature, can lead misfolding aggregation, consequently, dysfunction (2van Oosten-Hawle P. Morimoto R.I. Organismal proteostasis: Role cell-nonautonomous regulation transcellular chaperone signaling.Genes Dev. 2014; 28: 1533-1543Crossref Scopus (60) sense, rapidly respond, adapt new for survival. Organisms bacteria mammals evolved similar varying responses successfully. Some these strategies include modulations cascades, transcriptional programs, posttranslational modifications, dynamic assembly RNA condensates (ribonucleoprotein [RNP] granules) through liquid–liquid phase separation Scholar, 3de la Fuente M. Valera S. Martínez-Guitarte J.L. ncRNAs thermoregulation: A view prokaryotes eukaryotes.FEBS Lett. 2012; 586: 4061-4069Crossref (0) 4Protter D.S.W. Parker R. Principles properties granules.Trends Cell 2016; 26: 668-679Abstract Full Text PDF (621) 5Pohl C. Dikic I. Cellular control ubiquitin-proteasome system autophagy.Science. 2019; 366: 818-822Crossref (267) 6Pomatto L.C.D. Davies K.J.A. The role declining adaptive ageing.J. Physiol. 2017; 595: 7275-7309Crossref (76) 7Yasuda Tsuchiya H. Kaiho A. Guo Q. Ikeuchi K. Endo Arai N. Ohtake F. Murata Inada T. Baumeister W. Fernández-Busnadiego Tanaka Saeki Y. Stress- ubiquitylation-dependent proteasome.Nature. 2020; 578: 296-300Crossref (84) Several converge sustain sudden acute conditions. Increases temperature universal challenge encountered most organisms. historical reasons, thermal has been paradigm study response. Nowadays, studies additional relevance due increased exposure heatwaves derived climate change (8Tomanek L. importance physiological limits determining biogeographical range shifts global change: heat-shock response.Physiol. Biochem. Zool. 2008; 81: 709-717Crossref (114) 9Kassahn K.S. Crozier R.H. Pörtner H.O. Caley M.J. Animal performance stress: Responses tolerance at biological organisation.Biol. Rev. Camb. Philos. Soc. 2009; 84: 277-292Crossref (174) 10Sengupta Garrity Sensing temperature.Curr. 2013; 23: R304-R307Abstract (72) Increased energy result heat-induced denaturation thermally altered metabolic activity leading reactive oxygen species, all macromolecules, including (11Somero G.N. temperature: Function, regulation, evolution.J. Exp. Zool Ecol. Integr. 333: 379-397Crossref (44) Cells load unfolded modulating chaperones, also (HSPs) (12Parsell D.A. Lindquist tolerance: Degradation reactivation damaged proteins.Annu. Genet. 1993; 27: 437-496Crossref 13Kültz Molecular evolutionary basis OF response.Annu. 2005; 67: 225-257Crossref (977) 14Daugaard Rohde Jäättelä 70 family: Highly homologous overlapping distinct functions.FEBS 2007; 581: 3702-3710Crossref (830) 15Rosenzweig Nillegoda N.B. Mayer M.P. Bukau B. Hsp70 network.Nat. Mol. 20: 665-680Crossref (295) (HSR) refers HSPs, it common widely studied stress. play central lifecycle because promote folding nascent polypeptides into native/functional configurations prevent aggregation 16Morimoto response: Systems biology proteotoxic aging disease.Cold Spring Harb. Symp. Quant 2011; 76: 91-99Crossref (263) collaborate mechanisms, system, autophagy, target aggregates whose native state cannot recovered degradation (5Pohl 17Wang Le W.D. Autophagy system.Adv. Med. 1206: 527-550Crossref (37) Given are network, undertake several adaptations favor synthesis expense decreasing (Fig. 1). Biochemical approaches highlight unique HSP expression. spatiotemporal resolution precise now being uncovered using high-resolution quantitative microscopy. Gene adaptions during together protect macromolecules promptly cytoplasmic nuclear activities once permissive restored (3de coordinates other protective like formation RNP integrated (ISR) repress translation initiation. ISR HSR actions endoplasmic reticulum (ER) mitochondria preserve across compartments. ranging plants genes encoding HSPs. grouped families based on apparent weight (18Jayaraj G.G. Hipp M.S. Hartl F.U. Functional modules network.Cold Perspect. 12a033951Crossref (47) 19Kampinga H.H. Hageman J. Vos Kubota Tanguay R.M. Bruford E.A. Cheetham M.E. Chen Hightower L.E. Guidelines nomenclature human proteins.Cell Stress Chaperones. 14: 105-111Crossref (813) HSP70 HSP90 functionally relevant (15Rosenzweig 20Abisambra J.F. Blair L.J. Hill S.E. Jones J.R. Kraft Rogers Koren Jinwal U.K. Lawson Johnson A.G. Wilcock O'Leary J.C. Jansen-West Muschol Golde T.E. et al.Phosphorylation dynamics regulate Hsp27-mediated rescue neuronal plasticity deficits tau transgenic mice.J. Neurosci. 2010; 30: 15374-15382Crossref (78) They ATP-dependent cooperate small HSP110. Cochaperones J-domain family modulate accelerating ATP hydrolysis, participating substrate recognition refolding 2) (21Gamerdinger Hajieva Kaya A.M. Wolfrum U. Behl Protein involves recruitment macroautophagy pathway BAG3.EMBO 889-901Crossref (385) 22Kumar Ambasta R.K. Veereshwarayya V. Rosen K.M. Kosik Band Mestril Patterson Querfurth H.W. CHIP interact beta-APP proteasome-dependent manner influence Abeta metabolism.Hum. 16: 848-864Crossref 23Lindquist 1986; 55: 1151-1191Crossref 24Lindquist Craig 1988; 22: 631-677Crossref 25Liu Liang Zhou Structural analysis Hsp70/Hsp40 system.Protein Sci. 29: 378-390Crossref (28) 26Petrucelli Dickson Kehoe Taylor Snyder Grover De Lucia McGowan E. Lewis Prihar Kim Dillmann W.H. Browne Hall Voellmy al.CHIP ubiquitination, aggregation.Hum. 2004; 13: 703-714Crossref (560) 27Ritossa puffing pattern induced DNP drosophila.Experientia. 1962; 18: 571-573Crossref (1424) further categorized constitutive inducible steady-state levels. some upregulated extent upon Among them, fastest (23Lindquist Interestingly, highly conserved among species having amino acid similarity 50% Homo sapiens Escherichia coli, while domains 96% similar, highlights its vital adaption changing (28Sørensen J.G. Kristensen T.N. Loeschcke ecological proteins: Heat proteins.Ecol. 2003; 6: 1025-1037Crossref Scholar).Figure 2The HSC70/HSP70 retaining proteostasis. illustration depicts significant tasks network inside (Starting top left tile) Under nonstress HSC70 provides cotranslational polypeptide obtain conformation; helps proteins; transports cytoplasm where assisted mitochondrial (mtHSP70) HSP60 attain involved complex and/or disassembly; leads lysosome chaperone-mediated autophagy (236Massey Kiffin Cuervo Pathophysiology autophagy.Int. 36: 2420-2434Crossref (148) 237Majeski A.E. Dice Mechanisms 2435-2444Crossref (299) (Continuing bottom During lack exit ribosome tunnel represses elongation stage. resolves granules so sequestered mRNAs recovery stress; targets terminally proteasomal degradation; autophagosome. HSP, protein.View Large Image Figure ViewerDownload Hi-res image Download (PPT) frame context undertaken eukaryotic temperature. We compare mounted suggest technological overcome gap our knowledge one main Their occurs downregulation constitutively expressed genes. Most short (around 2500 nucleotides) intronless, promoter contains more binding sites, elements (HSEs), association master factor 1 (HSF1) (29Brocchieri Conway de Macario A.J. hsp70 genome: Conservation differentiation patterns predict wide array specialized functions.BMC Evol. 8: 19Crossref (178) not expressed. However, loci neither present compact heterochromatin domain nor marked repressive epigenetic histone modification. 3′ end nucleosome-free body covered nucleosomes. bound paused polymerase II (RNAPII) (30Petesch S.J. Lis J.T. Rapid, transcription-independent loss nucleosomes over large chromatin loci.Cell. 134: 74-84Abstract (250) These characteristics stable repression facilitate prompt HSF1. HSF1 shuttles nucleus cytoplasm, kept inactive monomer members families. released trimerizes, localizes binds HSE, comprised least three nGAAn repeats organized head tail promoters products (31Anckar Sistonen Regulation Implications disease.Annu. 80: 1089-1115Crossref (462) 32Vihervaara glance.J. 127: 261-266Crossref (169) Scholar) 3). domains, oligomerization next N terminus, trans-activation C terminus induces initiation elongation, regulatory middle negatively regulates By forming trimer, affinity HSE increases each trimer repeat domain. sufficient accompanied extensive modifications. undergoes hyperphosphorylation serine threonine residues cover up 90% (33Björk J.K. mammalian family.FEBS 277: 4126-4139Crossref 34Gomez-Pastor Burchfiel E.T. Thiele D.J. factors roles physiology disease.Nat. 2018; 19: 4-19Crossref (272) 35Guettouche Boellmann Lane W.S. Analysis phosphorylation experiencing stress.BMC 4Crossref (222) 36Nakai HSF regulation.Nat. Struct. 93-95Crossref (15) only few serines 230 326, necessary (35Guettouche 37Boellmann Guettouche Fenna Mnayer DAXX interacts enhances activity.Proc. Natl. Acad. 101: 4100-4105Crossref (70) Concomitantly, sumo groups inhibitory effect removed (38Hietakangas Anckar Blomster H.A. Fujimoto Palvimo J.J. Nakai PDSM, motif phosphorylation-dependent SUMO modification.Proc. 2006; 103: 45-50Crossref (374) acetylation lysines 116 118 favors activity, whereas lysine localization Acetylation hours after decrease (39Westerheide S.D. Stevens S.M. Stress-inducible deacetylase SIRT1.Science. 323: 1063-1066Crossref (530) summary, regulated under various stresses. Although modifications identified, many others, well responsible remains elucidated. combination titration demonstrated Recent work yeast, Saccharomyces cerevisiae, allowed building simple mathematical model points dissociation HSP70/HSP90 first "switch on" step feedback switch off (40Krakowiak Zheng X. Patel Feder Z.A. Anandhakumar Valerius Gross D.S. Khalil A.S. Pincus Hsf1 constitute two-component loop response.Elife. 7e31668Crossref (33) 41Zheng Krakowiak Beyzavi Ezike Dynamic phosphorylation.Elife. 5e18638Crossref (97) al. (41Zheng identified sites were able phosphorylations no but instead favoring mediator complex. Additionally, eEF1A noncoding HSR1 activating form nucleoprotein stimulate trimerization (42Shamovsky Ivannikov Kandel E.S. Gershon Nudler RNA-mediated cells.Nature. 440: 556-560Crossref (271) Following shock, recruits multiple cofactors (43Chen Yu Yang Temple Harbinski Gao Wilson Pagliarini Identification mixed lineage leukemia 1(MLL1) coactivator 1(HSF1) 90 (HSP90) inhibition.J. Chem. 289: 18914-18927Abstract 44Jonkers Getting speed II.Nat. 2015; 167-177Crossref (468) 45Mason P.B. Cooperative competitive interactions promoter.J. 1997; 272: 33227-33233Abstract 46Park J.M. Werner Y.J. Mediator, holoenzyme, directly recruited shock.Mol. 2001; 9-19Abstract (113) Scholar), SGO2, subunit MED12, essential strong (47Takii Matsumoto Srivastava Katiyar Nakayama K.I. pericentromeric shugoshin 2 cooperates Pol recruitment.EMBO 38e102566Crossref (11) SGO2 hypophosphorylated RNAPII Transcription then P-TEFb, mediated (48Lis Mason Peng Price D.H. P-TEFb kinase loci.Genes 792-803Crossref induce C-terminal RNAPII, 49Marshall N.F. Xie Z. Control potential novel Carboxyl-terminal kinase∗.J. 1996; 271: 27176-27183Abstract positioned along removed. remodelers SWI/SNF FACT Spt6 Drosophila melanogaster (D. melanogaster) within minutes remove Besides HSF1, depends relocation membrane speckles (50Khanna Hu Belmont transgene directed motion facilitates activation.Curr. 24: 1138-1144Abstract (85) 51Vera Singer regulation: jumps shocked.Curr. R396-R398Abstract rapid, active, unidirectional movement actin polymerization. locus sequence determines contain 2–phosphorylated components machinery, speckle yet identified. coalesce discrete spots stimulation. interallelic clustering interaction HSP104 HSP12 transcription. suggested presence factories formed could coregulated (52Chowdhary Kainth undergo alteration three-dimensional structure genome organization stress.Mol. 37e00292-17Crossref (17) upregulation non-HSP cytoskeleton oxidative massive thousands (For review: (53Vihervaara Duarte F.M. driving responses.Nat. 385-397Crossref (98) Scholar)). Detailed position polymerases, landscape explain preferences (54Mueller Mieczkowski Kundu Wang Sadreyev Tolstorukov M.Y. Kingston R.E. Widespread nucleosome accessibility without occupancy induction.Genes 31: 451-462Crossref (55) 55Vihervaara Mahat D.B. Guertin Chu Danko C.G. Transcriptional pre-wired enhancer architecture.Nat. Commun. 255Crossref (65) does remodeling topology associated S2 (56Ray Munn P.R. Vihervaara Ozer Chromatin conformation cha

Language: Английский

Citations

104
eIF4F is a thermo-sensing regulatory node in the translational heat shock response DOI Creative Commons
Christine Desroches Altamirano,

Moo-Koo Kang,

Mareike A. Jordan

et al.

Molecular Cell, Journal Year: 2024, Volume and Issue: 84(9), P. 1727 - 1741.e12

Published: March 27, 2024

Heat-shocked cells prioritize the translation of heat shock (HS) mRNAs, but underlying mechanism is unclear. We report that HS in budding yeast induces disassembly eIF4F complex, where eIF4G and eIF4E assemble into translationally arrested mRNA ribonucleoprotein particles (mRNPs) granules (HSGs), whereas eIF4A promotes translation. Using vitro reconstitution biochemistry, we show a conformational rearrangement thermo-sensing eIF4A-binding domain dissociates assembly with HS-mRNPs, which recruit additional factors, including Pab1p eIF4E, to form multi-component condensates. extracts cellular experiments, demonstrate HS-mRNPs condensates repress associated deplete factors are required for housekeeping translation, mRNAs can be efficiently translated by eIF4A. conclude complex node regulates during HS.

Language: Английский

Citations

20
Hsp90‐mediated regulation of DYRK3 couples stress granule disassembly and growth via mTORC1 signaling DOI Creative Commons
Laura Mediani, Francesco Antoniani,

Veronica Galli

et al.

EMBO Reports, Journal Year: 2021, Volume and Issue: 22(5)

Published: March 19, 2021

Article19 March 2021Open Access Source DataTransparent process Hsp90-mediated regulation of DYRK3 couples stress granule disassembly and growth via mTORC1 signaling Laura Mediani Department Biomedical, Metabolic Neural Sciences, Centre for Neuroscience Nanotechnology, University Modena Reggio Emilia, Modena, Italy Search more papers by this author Francesco Antoniani ItalyThese authors are contributed equally to work as second, third Veronica Galli Jonathan Vinet Genomic Post-Genomic Center, IRCCS Mondino Foundation, Pavia, Arianna Dorotea Carrà Ilaria Bigi Vadreenath Tripathy Center Regenerative Therapies TU Dresden, Technische Universität Germany Tatiana Tiago Marco Cimino Giuseppina Leo Triana Amen orcid.org/0000-0003-4808-7806 Experimental Neurodegeneration, Medical Göttingen, Daniel Kaganovich orcid.org/0000-0003-2398-1596 Cristina Cereda orcid.org/0000-0001-9571-0862 Orietta Pansarasa Jessica Mandrioli Neuroscience, St. Agostino Estense Hospital, Azienda Ospedaliero Universitaria di Priyanka Tripathi Institute Neuropathology, RWTH Aachen Aachen, Dirk Troost (Neuro)Pathology, Amsterdam UMC, Amsterdam, The Netherlands Eleonora Aronica Johannes Buchner orcid.org/0000-0003-1282-7737 Integrated Protein Science Munich at the Chemie, München, Garching, Anand Goswami Jared Sterneckert Simon Alberti Corresponding Author [email protected] orcid.org/0000-0003-4017-6505 Biotechnology (BIOTEC), Molecular Cellular Bioengineering (CMCB), Serena Carra orcid.org/0000-0003-0939-0140 Information Mediani1, Antoniani1, Galli1, Vinet1,2, Carrà1, Bigi1, Tripathy3, Tiago1, Cimino1, Leo1, Amen4, Kaganovich4, Cereda2, Pansarasa2, Mandrioli5, Tripathi6, Troost7, Aronica7, Buchner8, Goswami6, Sterneckert3, *,9 *,1 1Department 2Genomic 3Center 4Department 5Department 6Institute 7Department 8Center 9Biotechnology *Corresponding author. Tel: +49 351 46340243; E-mail: (lead contact). +39 059 2055265; EMBO Reports (2021)22:e51740https://doi.org/10.15252/embr.202051740 PDFDownload PDF article text main figures. Peer ReviewDownload a summary editorial decision including letters, reviewer comments responses feedback. ToolsAdd favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Stress granules (SGs) dynamic condensates associated with protein misfolding diseases. They sequester stalled mRNAs factors, such subunit raptor, suggesting that SGs coordinate cell during after stress. However, molecular mechanisms linking SG dynamics remain undefined. We report chaperone Hsp90 is required dissolution. binds stabilizes dual-specificity tyrosine-phosphorylation-regulated kinase 3 (DYRK3) in cytosol. Upon inhibition, dissociates from becomes inactive. Inactive subjected two different fates: it either partitions into SGs, where protected irreversible aggregation, or degraded. In presence Hsp90, active promotes disassembly, restoring translation. Thus, links adaptation regulating activity key involved condensate translation restoration. SYNOPSIS other part binding stabilizing (DYRK3). Inhibition loss results destabilization, persistence failure restore regulates stability activity. Partitioning protects aggregation. mTORC1-dependent DYRK3. ALS fibroblasts motor neurons show reduced expression Introduction ribonucleoprotein assemblies form response conditions transiently translationally along 40S ribosomal subunits, enzymes, molecules, p90 S6 raptor (Kedersha et al, 2013). Raptor-mediated sequestration inside cells apoptosis (Thedieck relief, dissolution, subsequent release reactivation complex (Wippich This suggests hubs metabolism changes environmental conditions, thus enabling stressors details how cellular assembly dissolution have so far eluded us. Recent impaired dysfunction death. dysfunctionality emerging an important pathomechanism number age-related neurodegenerative diseases amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTD), Alzheimer disease (AD). these diseases, disassemble has been linked disturbances RNA development aggregates cause neuronal death (Zhang 2019). Hence, identifying characterizing factors govern will provide targets therapeutic potential treat uncurable Surprisingly, although we gained detailed knowledge on promote 1999; Matsuki 2013; Van Treeck 2018), still poorly understood. RNA-binding proteins (RBPs) high concentrations free crucial drive (Bounedjah 2012; Ciryam 2015; Kroschwald 2018; Guillen-Boixet 2020; Sanders Yang 2020). many RBPs assemble propensity misfold aggregate, which can affect material properties their ability dissolve (Molliex Patel 2015). Such aberrant also arise accumulation misfolding-prone proteins, defective products (DRiPs) ALS-linked C9orf72 dipeptide repeat (Ganassi 2016; Lee Mateju 2017; As result, under close surveillance quality control (PQC) system. PQC system includes chaperones, recognize aberrantly folded refold misfolded well ubiquitin–proteasome autophagy degradation systems, clear (Balchin 2016). Indeed, chaperones Hsp70 valosin-containing (VCP) able prevent maintaining 2017). addition, SGs. When machinery fails, persisting targeted proteasomal involving p62/SQSTM1 ZFAND1/VCP (Buchan Chitiprolu Turakhiya 2018). our understanding its infancy do not know whether maintain healthy state. One essential ubiquitous folding maturation large variety components, (Jain Markmiller suggested regulate processing bodies (PBs) (Matsumoto 2011), cytosolic RNA-protein mRNA storage (Standart Weil, Moreover, promotes, through yet unknown mechanism, recruitment argonaute 2, eIF4E, partner eIF4E transporter (4E-T) (Pare 2009; Suzuki 2009). agreement this, recent proteomic analysis fungal pathogen identified novel role PB components (O'Meara despite well-known function tolerance, unclear functionality. Here, use genetics fluorescence microscopy fixed live study functional effects mammalian cells. Results delays Hsp70-independent manner cells, majority recovery phase, only minor fraction (< 5–10%) clearance Using time-lapse HeLa-Kyoto expressing marker G3BP2-GFP, confirmed most sodium arsenite induced when removed; dissolved regardless lysosomal proteases were inactivated ammonium chloride (Fig EV1A Movie EV1), does play major role. By contrast, inhibition ATPase VER-155008 resulted (VER; Figs 1A EV1B, literature was absolute: could inhibitor, slower kinetics persisted ca. 20–30% 4 h removal. Combined data suggest regulated additional besides Hsp70. Click here expand figure. Figure EV1. independently Related Fig 1 A. Kinetics living absence chloride. G3BP2-GFP treated (50 µM) 45 min. Then, allowed recover drug-free medium (recovery control) (NH4Cl, 20 mM). Images taken over time period every 10 Dashed lines = 95% confidence intervals. Number counted: 193 (Recovery Control); 104 B. left untreated (control) GA (5 µM), 17AAG VER (40 h. Cells fixed, percentage counted. n independent experiments, ± s.e.m.; 126–185 counted/sample. n.s.: non-significant (one-way ANOVA). C, D. HEK293 express SG-resident PABP endogenously tagged fluorescent probe Dendra2 (PABPC1-Dendra2). PABPC1-Dendra2 HEK293K (100 mM), µM). 299 control); 160 mM); 361 (GA 5 µM); (17AAG 259 (VER 40 E. Confocal showing colocalize DRiPs. HeLa OP-puro (25 DRiPs visualized click chemistry, while Hsc70, Hsp70, α, β immunostaining. Scale bar µm. F. Quantitation DRiP enrichment Automated imaging segmentation based G3BP signal. Data presented histogram. HS 43.5°C alone segmented: 2,198 (control); 3,056 (VER); 1,484 (GA); 3,220 (17AAG); P < 10−10 G, H. stably V5-tagged inducible tetracycline (Flp-In) cultured (V5-HSP70 OFF V5-HSP70 ON, respectively). (G) extracts prepared 24 treatment, levels analyzed immunoblotting. TUBA4A used loading control. (H) (500 min, followed 90 then 115-378 counted/sample; I. h, respectively. stained endogenous DCP1A, P-bodies. Nucleic acid DAPI. P-bodies treatment shown. h: 104–124 counted/sample, 0.02; 111–178 10−7 J. lipofected cDNA encoding mRFP-DCP1A. post-transfection, 2 Representative images GFP-G3BP2 mRFP-DCP1A 0, 30, 60, 90, 120 min Download figure PowerPoint 1. types A, inhibitors. Control) µM 0.5 Control, A); 340 187 209 363 B); 125 353 1.5 G3BP1-mCherry 72 non-targeting siRNA siRNAs specific α β. (C) Efficacy knockdown HSPA1A verified total extracts. (D) medium. reported. 159 (siRNA 166 + β). 37°C TIA-1, 605 510 VER); 1,334 GA); 468 17AAG). 3–4 s.e.m. 0.01 (One-way MG132 (20 439 649 637 0.0001 information: EV1 Movies EV1–EV5. (Taipale 2010; Biebl Buchner, 2019; Moran Luengo 2019) interacts (Markmiller Yet, currently dynamics. To test idea, measured well-established inhibitors activity, geldanamycin (GA) 17-Allylamino-17-demethoxygeldanamycin (17AAG) (Schulte Neckers, 1998). experiment, determined induces spontaneous assembly. up 3% EV1B). added phase both delayed B EV2), dose-dependent effect 1B EV2). Delayed upon be reproduced line EV1C D, EV3 EV4, HEK293T cells) observed siRNA-mediated 1C D EV5). Of note, stress, heat shock proteasome inhibitor 1E F). regulator type line. (or persistence) accumulate example, dysfunctional asked affects Although assisting subset newly synthesized (Schopf 2017), did observe colocalization alpha beta EV1E). contrast HSPA8 EV1E), targeting (Hartl Hayer-Hartl, 2002; Ganassi VER, 17AAG, GA, quantified microscopy. caused strong EV1F; 2016), led milder EV1F). Next, if upregulation assists prevents rescue found induction EV1G) EV1H). Together, demonstrate than Short-term It shown long-term decreases (Suzuki PBs cytoplasmic mRNP present growing increase size 2005). intimately connected SGs: equilibrium, share several interact, allowing transfer mRNPs between may perturb equilibrium PBs, indirectly affecting hypothesis, first using antibody PB-resident DCP1A. previous findings, disappearance EV1I), whereas had no EV1I). Tre

Language: Английский

Citations

61
Adaptive preservation of orphan ribosomal proteins in chaperone-dispersed condensates DOI
Asif Ali, Rania Garde, Olivia Schaffer

et al.

Nature Cell Biology, Journal Year: 2023, Volume and Issue: 25(11), P. 1691 - 1703

Published: Oct. 16, 2023

Language: Английский

Citations

35
Transcriptome-wide mRNA condensation precedes stress granule formation and excludes stress-induced transcripts DOI Creative Commons
Hendrik Glauninger, Jared A.M. Bard, Caitlin J. Wong Hickernell

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 16, 2024

Abstract Stress-induced condensation of mRNA and proteins into stress granules is conserved across eukaryotes, yet the function, formation mechanisms, relation to well-studied transcriptional responses remain largely unresolved. exposure ribosome-free following translational shutoff thought cause by allowing new multivalent RNA-dependent interactions, with RNA length associated interaction capacity driving increased condensation. Here we show that, in striking contrast, virtually all species condense response multiple unrelated stresses budding yeast, plays a minor role, instead, stress-induced transcripts are preferentially excluded from condensates, enabling their selective translation. Using both endogenous genes reporter constructs, that translation initiation blockade, rather than resulting RNA, causes These initiation-inhibited condensates (TIICs) biochemically detectable even when granules, defined as microscopically visible foci, absent or blocked. TIICs occur unstressed yeast cells, and, during stress, grow before appearance granules. primarily due timing expression, sequence features. Together, our results reveal simple system which cells redirect activity newly synthesized broad implications for cellular regulation changing conditions.

Language: Английский

Citations

12
The Heat Shock Response as a Condensate Cascade DOI
Annisa Dea, David Pincus

Journal of Molecular Biology, Journal Year: 2024, Volume and Issue: 436(14), P. 168642 - 168642

Published: June 5, 2024

Language: Английский

Citations

11
The life and death of RNA across temperatures DOI Creative Commons
Attila Becskei, Sayanur Rahaman

Computational and Structural Biotechnology Journal, Journal Year: 2022, Volume and Issue: 20, P. 4325 - 4336

Published: Jan. 1, 2022

Temperature is an environmental condition that has a pervasive effect on cells along with all the molecules and reactions in them. The mechanisms by which prototypical RNA sense withstand heat have been identified mostly bacteria archaea. relevance of these phenomena is, however, broader, similar recently found throughout tree life, from sex determination reptiles to adaptation viral polymerases, genetic disorders humans. We illustrate temperature dependence metabolism examples synthesis degradation mRNAs, review emerged questions. Are exposed greater variations gradients than previously surmised? How do reconcile conflicting thermal stability requirements primary tertiary structures RNAs? To what extent enzymes contribute compensation reaction rates mRNA turnover lowering energy barrier catalyzed reactions? conclude ecological, forensic applications temperature-dependence biotechnological aspects vaccine production.

Language: Английский

Citations

35
Fungal survival under temperature stress: a proteomic perspective DOI Creative Commons
Nurlizah Abu Bakar, Saiful Anuar Karsani, Siti Aisyah Alias

et al.

PeerJ, Journal Year: 2020, Volume and Issue: 8, P. e10423 - e10423

Published: Dec. 15, 2020

Background Increases in knowledge of climate change generally, and its impact on agricultural industries specifically, have led to a greater research effort aimed at improving understanding the role fungi various fields. Fungi play key soil ecosystems as primary agent decomposition, recycling organic nutrients. also include important pathogens plants, insects, bacteria, domestic animals humans, thus highlighting their importance many contexts. Temperature directly affects fungal growth protein dynamics, which ultimately will cascade through affect crop performance. To study changes global complement fungi, proteomic approaches been used examine links between temperature stress profiles. Survey methodology objectives A traditional rather than systematic review approach was taken focus responses elucidated using approaches. The effects metabolic pathways and, particular, heat shock proteins (HSPs) are discussed. objective this is provide an overview proteomes. Concluding remarks Elucidating response under useful context increasing sensitivity resilience challenges posed by contemporary processes. Although useful, more thorough work needed such combining data from multiple -omics platforms order develop deeper factor influencing controlling cell physiology. This information can be beneficial identify potential biomarkers for monitoring environmental soil, including vital human society economy.

Language: Английский

Citations

44
The UBA domain of conjugating enzyme Ubc1/Ube2K facilitates assembly of K48/K63‐branched ubiquitin chains DOI Creative Commons
Lukas Pluska, Ernst Jarosch, Henrik Zauber

et al.

The EMBO Journal, Journal Year: 2021, Volume and Issue: 40(6)

Published: Feb. 12, 2021

The assembly of a specific polymeric ubiquitin chain on target protein is key event in the regulation numerous cellular processes.Yet, mechanisms that govern selective synthesis particular polyubiquitin signals remain enigmatic.The homologous ubiquitin-conjugating (E2) enzymes Ubc1 (budding yeast) and Ube2K (mammals) exclusively generate linked through lysine 48 (K48).Uniquely among E2 enzymes, harbor ubiquitin-binding UBA domain with unknown function.We found this preferentially interacts chains 63 (K63).Based structural modeling, vitro ubiquitination experiments, NMR studies, we propose aligns K63linked facilitates K48/ K63-branched conjugates.Genetic proteomics experiments link activity domain, hence formation unusual topology, to maintenance proteostasis.

Language: Английский

Citations

39