Reversible phase separation of HSF1 is required for an acute transcriptional response during heat shock

Zhang, Hongchen; Shao, Shuang; Zeng, Yong; Wang, Xiaotian; Qin, Yizhi; Ren, Qiunan; Xiang, ShengQi; Wang, Yuxin; Xiao, Junyu; Sun, Yujie

doi:10.1038/s41556-022-00846-7

Cited by 96 publications

(73 citation statements)

References 60 publications

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“…The linearity of the calibrated localization number with molecule density provides access to quantitative analysis. For example, to determine the density change of target molecule under different treatment using STORM imaging, the change of molecule density usually reflected by localization number of the target molecule 29 . Statistical techniques such as two-sample t-test can be used to determine whether two groups of density are equal.…”

Section: Resultsmentioning

confidence: 99%

Identification and calibration of ultrabright localizations to eliminate quantification error in SMLM

Chen

Niu

et al. 2022

Preprint

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Single molecule localization microscopy (SMLM) is irreplaceable among super-resolution microscopies in revealing biological ultra-structures, given its unmatched high resolution. However, its sub-optimal quantitative capability, which is critical for characterizing true biomolecular organization of ultra-structures in cells, has hindered its widest application in biomedical research. Here, in SMLM imaging of cellular structures such as lipid rafts and microtubules with saturation labelling, we identified ultra-bright localizations, each of which is contributed by simultaneous emission of multiple molecules within a diffraction-limit region and has been regarded before as a regular localization from single molecule. Consistently, ultra-bright localizations are also observed in simulated SMLM imaging of endoplasmic reticulum or microtubules from public resource. Furthermore, after calibrating each ultrabright localization into multiple single-molecule localizations using the photon-number-based models, the density of total localizations shows linear correlation with the true molecule density, presenting SMLM with new reconstruction method as a quantitative analysis approach. Therefore, identification and dissection of ultra-bright localizations in SMLM enable the close and quantitative estimate of the true biomolecular organization.

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Section: Resultsmentioning

confidence: 99%

Identification and calibration of ultrabright localizations to eliminate quantification error in SMLM

Chen

Niu

et al. 2022

Preprint

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“…Homogeneous oligomers of C233 would dynamically move between the cytoplasm and the condensates. Protein condensates are formed as a result of stress such as heat shock and aging (Jawerth et al , 2020; Zhang et al , 2022). The biology of condensates is likely various because the condensates formed in response to acute stress such as heat shock are reversible (Alagar Boopathy et al , 2022); however, these processes could be perturbed in the presence of disease proteins and/or during ageing (Shin & Brangwynne, 2017).…”

Section: Discussionmentioning

confidence: 99%

Hetero-oligomerization of TDP-43 carboxy-terminal fragments with cellular proteins contributes to proteotoxicity

Kitamura

Fujimoto

Kawashima

et al. 2022

Preprint

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Carboxy terminal fragments (CTFs) of TDP-43 contain an intrinsically disordered region (IDR) and form cytoplasmic condensates containing amyloid fibrils. Such condensates are toxic and associated with pathogenicity in several neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. However, the molecular details of how the domain of TDP-43 CTFs leads to condensation and cytotoxicity remain elusive. Here, we show that truncated RNA/DNA-recognition motif (RRM) at the N-terminus of TDP-43 CTFs is assembled and leads to the structural transition of the IDR, whereas the IDR itself of TDP-43 CTFs is difficult to assemble even if they are proximate intermolecularly. Hetero-oligomers of TDP-43 CTFs that have recruited other proteins that are essential for proteostasis into low mobile condensates are more toxic than homo-oligomers inside highly mobile condensates, implicating loss-of-function of the endogenous proteins by such oligomers, not necessarily the condensates, is associated with cytotoxicity. Furthermore, such toxicity of TDP-43 CTFs was cell-nonautonomously affected in the nematodes. We speculate that the misfolding and oligomeric characteristics of the truncated RRM at the N-terminus of TDP-43 CTFs define their condensation properties and toxicity by implanting and transmitting structures with toxic properties of the truncated RRM into the IDR.

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“…In mammalian cells, the resolution of nSBs correlates with HSF1 activity, transcription of hsp target genes, and cell survival at the single cell level ( Gaglia et al, 2020 ). A recent mammalian study discovered the existence of both HSF1 nSBs at non- hsp gene loci and smaller HSF1 condensates at transcriptionally active hsp gene loci upon heat shock ( Zhang et al, 2022 ). In this study, HSF1 in the condensates was found to colocalize with transcription apparatus factors, including RNA Pol II, BRD4, MED1, and CYCT1 ( Zhang et al, 2022 ).…”

Section: Discussionmentioning

confidence: 99%

The CBP-1/p300 Lysine Acetyltransferase Regulates the Heat Shock Response in C. elegans

Barrett

2022

Front. Aging

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The decline of proteostasis is a hallmark of aging that is, in part, affected by the dysregulation of the heat shock response (HSR), a highly conserved cellular response to proteotoxic stress in the cell. The heat shock transcription factor HSF-1 is well-studied as a key regulator of proteostasis, but mechanisms that could be used to modulate HSF-1 function to enhance proteostasis during aging are largely unknown. In this study, we examined lysine acetyltransferase regulation of the HSR and HSF-1 in C. elegans. We performed an RNA interference screen of lysine acetyltransferases and examined mRNA expression of the heat-shock inducible gene hsp-16.2, a widely used marker for HSR activation. From this screen, we identified one acetyltransferase, CBP-1, the C. elegans homolog of mammalian CREB-binding protein CBP/p300, as a negative regulator of the HSR. We found that while knockdown of CBP-1 decreases the overall lifespan of the worm, it also enhances heat shock protein production upon heat shock and increases thermotolerance of the worm in an HSF-1 dependent manner. Similarly, we examined a hallmark of HSF-1 activation, the formation of nuclear stress bodies (nSBs). In analyzing the recovery rate of nSBs, we found that knockdown of CBP-1 enhanced the recovery and resolution of nSBs after stress. Collectively, our studies demonstrate a role of CBP-1 as a negative regulator of HSF-1 activity and its physiological effects at the organismal level upon stress.

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Reversible phase separation of HSF1 is required for an acute transcriptional response during heat shock

Cited by 96 publications

References 60 publications

Identification and calibration of ultrabright localizations to eliminate quantification error in SMLM

Identification and calibration of ultrabright localizations to eliminate quantification error in SMLM

Hetero-oligomerization of TDP-43 carboxy-terminal fragments with cellular proteins contributes to proteotoxicity

The CBP-1/p300 Lysine Acetyltransferase Regulates the Heat Shock Response in C. elegans

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