Regulation of the heat shock response by thiol-reactive compounds in the yeast Saccharomyces cerevisiae

Cells govern their activities and modulate their interactions with the environment to achieve homeostasis. The heat shock response (HSR) is one of the most well studied fundamental cellular responses to environmental and physiological challenges, resulting in rapid synthesis of heat shock proteins (HSPs), which serve to protect cellular constituents from the deleterious effects of stress. In addition to its role in cytoprotection, the HSR also influences lifespan and is associated with a variety of human diseases including cancer, aging and neurodegenerative disorders. In most eukaryotes, the HSR is primarily mediated by the highly conserved transcription factor HSF1, which recognizes target hsp genes by binding to heat shock elements (HSEs) in their promoters. In recent years, significant efforts have been made to identify small molecules as potential pharmacological activators of HSF1 that could be used for therapeutic benefit in the treatment of human diseases relevant to protein conformation. However, the detailed mechanisms through which these molecules drive HSR activation remain unclear. In this work, I utilized the baker's yeast Saccharomyces cerevisiae as a model system to identify a group of thiol-reactive molecules including oxidants, transition metals and metalloids, and electrophiles, as potent activators of yeast Hsf1. Using an artificial HSE-lacZ reporter and the glucocorticoid receptor system (GR), these diverse thiol-reactive compounds are shown to activate Hsf1 and inhibit Hsp90 chaperone complex activity in a reciprocal, dose-dependent manner. To further understand whether cells sense these reactive compounds through accumulation of unfolded proteins, the proline analog azetidine-2-carboxylic acid (AZC) and protein cross-linker dithiobis(succinimidyl propionate) (DSP) were used to force misfolding of nascent polypeptides and existing cytosolic proteins, respectively. Both unfolding reagents display kinetic HSP induction profiles dissimilar to those generated by thiol-reactive compounds. Moreover, AZC treatment leads to significant cytotoxicity, which is not observed in the presence of the thiol-reactive compounds at the concentrations sufficient to induce Hsf1. Additionally, DSP treatment has little to no effect on Hsp90 functions. Together with the ultracentrifugation analysis of cell lysates that detected no insoluble protein aggregates, my data suggest that at concentrations sufficient to induce Hsf1, thiol-reactive compounds do not induce the HSR via a mechanism based on accumulation of unfolded cytosolic proteins. Another possibility is that thiol-reactive compounds may influence aspects of the protein quality control system such as the ubiquitin-proteasome system (UPS). To address this hypothesis, β-galactosidase reporter fusions were used as model substrates to demonstrate that thiol-reactive compounds do not inhibit ubiquitin activating enzymes (E1) or proteasome activity. Therefore, thiol-reactive compounds do not activate the HSR by inhibiting UPS-dependent protein degradation. I therefore hypothesized that these molecules may directly inactivate protein chaperones, known as repressors of Hsf1. To address this possibility, a thiol-reactive biotin probe was used to demonstrate in vitro that the yeast cytosolic Hsp70 Ssa1, which partners with Hsp90 to repress Hsf1, is specifically modified. Strikingly, mutation of conserved cysteine residues in Ssa1 renders cells insensitive to Hsf1 activation by cadmium and celastrol but not by heat shock. Conversely, substitution with the sulfinic acid and steric bulk mimic aspartic acid led to constitutive activation of Hsf1. Cysteine 303, located in the nucleotide-binding/ATPase domain of Ssa1, was shown to be modified in vivo by a model organic electrophile using Click chemistry technology, verifying that Ssa1 is a direct target for thiol-reactive compounds through adduct formation. Consistently, cadmium pretreatment promoted cells thermotolerance, which is abolished in cells carrying SSA1 cysteine mutant alleles. Taken together, these findings demonstrate that Hsp70 acts as a sensor to induce the cytoprotective heat shock response in response to environmental or endogenously produced thiol-reactive molecules and can discriminate between two distinct environmental stressors.

Susy Shock trans piradx : El inclasificable género colibrí

Mensualmente, en Buenos Aires, la artista trans Susy Shock presenta su arte performativo en Poemarios Trans Pirados y Noches Bizarras. Su reivindicación a lo monstruoso se inscribe en lo queer pues propone la imposibilidad de rotular, clasificar, o clausurar las derivas de deseo, género y sexo. Vislumbramos lo ?mascufemenino? en disputa: una deconstrucción de categorías naturalizadas que invita a repensarnos en cada escrito, canción y presentación. Su construcción de género deviene en "colibrí", recordando el Parakultural y al "clown-travesti-literario" Batato Barea. Así, advertimos la destrucción de binarismos en un cuerpo deseante y una militancia trans que transgrede la "matriz heterosexual"

Susy Shock trans piradx : El inclasificable género colibrí

Mensualmente, en Buenos Aires, la artista trans Susy Shock presenta su arte performativo en Poemarios Trans Pirados y Noches Bizarras. Su reivindicación a lo monstruoso se inscribe en lo queer pues propone la imposibilidad de rotular, clasificar, o clausurar las derivas de deseo, género y sexo. Vislumbramos lo ?mascufemenino? en disputa: una deconstrucción de categorías naturalizadas que invita a repensarnos en cada escrito, canción y presentación. Su construcción de género deviene en "colibrí", recordando el Parakultural y al "clown-travesti-literario" Batato Barea. Así, advertimos la destrucción de binarismos en un cuerpo deseante y una militancia trans que transgrede la "matriz heterosexual"

Susy Shock trans piradx : El inclasificable género colibrí

Mensualmente, en Buenos Aires, la artista trans Susy Shock presenta su arte performativo en Poemarios Trans Pirados y Noches Bizarras. Su reivindicación a lo monstruoso se inscribe en lo queer pues propone la imposibilidad de rotular, clasificar, o clausurar las derivas de deseo, género y sexo. Vislumbramos lo ?mascufemenino? en disputa: una deconstrucción de categorías naturalizadas que invita a repensarnos en cada escrito, canción y presentación. Su construcción de género deviene en "colibrí", recordando el Parakultural y al "clown-travesti-literario" Batato Barea. Así, advertimos la destrucción de binarismos en un cuerpo deseante y una militancia trans que transgrede la "matriz heterosexual"

Rapid acclimation of juvenile corals to CO2-mediated acidification by upregulation of heat shock protein and Bcl-2 genes

Corals play a key role in ocean ecosystems and carbonate balance, but their molecular response to ocean acidification remains unclear. The only previous whole-transcriptome study documented extensive disruption of gene expression, particularly of genes encoding skeletal organic matrix proteins, in juvenile corals (Acropora millepora) after short-term (3 d) exposure to elevated pCO2. In this study, whole-transcriptome analysis was used to compare the effects of such 'acute' (3 d) exposure to elevated pCO2 with a longer ('prolonged'; 9 d) period of exposure beginning immediately post-fertilization. Far fewer genes were differentially expressed under the 9-d treatment, and although the transcriptome data implied wholesale disruption of metabolism and calcification genes in the acute treatment experiment, expression of most genes was at control levels after prolonged treatment. There was little overlap between the genes responding to the acute and prolonged treatments, but heat shock proteins (HSPs) and heat shock factors (HSFs) were over-represented amongst the genes responding to both treatments. Amongst these was an HSP70 gene previously shown to be involved in acclimation to thermal stress in a field population of another acroporid coral. The most obvious feature of the molecular response in the 9-d treatment experiment was the upregulation of five distinct Bcl-2 family members, the majority predicted to be anti-apoptotic. This suggests that an important component of the longer term response to elevated CO2 is suppression of apoptosis. It therefore appears that juvenile A. millepora have the capacity to rapidly acclimate to elevated pCO2, a process mediated by upregulation of specific HSPs and a suite of Bcl-2 family members.

Heat shock resistance traits in specimens of the seaweed Gracilaria vermiculophylla originating from native and non-native populations

We compared the responses of native and non-native populations of the seaweed Gracilaria vermiculophylla to heat shock in common garden-type experiments. Specimens from six native populations in East Asia and from eight non-native populations in Europe and on the Mexican Pacific coast were acclimated to two sets of identical conditions before their resistance to heat shock was examined. The experiments were carried out twice - one time in the native range in Qingdao, China and one time in the invaded range in Kiel, Germany - to rule out effects of specific local conditions. In both testing sites the non-native populations survived heat shock significantly better than the native populations, The data underlying this statement are presented in https://doi.pangaea.de/10.1594/PANGAEA.859335. After three hours of heat shock G. vermiculophylla exhibited increased levels of heat shock protein 70 (HSP70) and of a specific isoform of haloperoxidase, suggesting that both enzymes could be required for heat shock stress management. However, the elevated resistance toward heat shock of non-native populations only correlated with an increased constitutive expression of heat shock protein 70 (HSP70). The haloperoxidase isoform was more prominent in native populations, suggesting that not only increased HSP70 expression, but also reduced allocation into haloperoxidase expression after heat shock was selected during the invasion history. The data describing expression of HSP70 and three different isoforms of haloperoxidase are presented in https://doi.pangaea.de/10.1594/PANGAEA.859358.

Combined effects of short-term ocean acidification and heat shock in a benthic copepod Tigriopus japonicus Mori

Warming of the world's oceans is predicted to have many negative effects on organisms as they have optimal thermal windows. In coastal waters, however, both temperatures and pCO2 (pH) exhibit diel variations, and biological performances are likely to be modulated by physical and chemical environmental changes. To understand how coastal zooplankton respond to the combined impacts of heat shock and increased pCO2, the benthic copepod Tigriopus japonicus were treated at temperatures of 24, 28, 32 and 36 °C to simulate natural coastal temperatures experienced in warming events, when acclimated in the short term to either ambient (LC, 390 µatm) or future CO2 (HC, 1000 µatm). HC and heat shock did not induce any mortality of T. japonicus, though respiration increased up to 32 °C before being depressed at 36 °C. Feeding rate peaked at 28 °C but did not differ between CO2 treatments. Expression of heat shock proteins (hsps mRNA) was positively related to temperature, with no significant differences between the CO2 concentrations. Nauplii production was not affected across all treatments. Our results demonstrate that T. japonicus responds more sensitively to heat shocks rather than to seawater acidification; however, ocean acidification may synergistically act with ocean warming to mediate the energy allocation of copepods.

Career Crisis? Impacts of Financial Shock on the Entry-Level Labor Market: Evidence from Thailand

We utilize Thailand's the financial crisis in 1997 as a natural experiment which exogenously shifts labor demand. Convincing evidence from the Thailand Labor Force Survey support the hypothesis that both employment opportunities and wages shrunk for new entrants after the crisis. We find that workers who entered before the crisis experienced job losses and wage losses. But these losses were smaller than those of new entrants after the crisis. We also find that new entrants after the crisis experienced a 10% reduction in the overtime wages compared to new entrants before the crisis.

Shock Transmission Mechanism of the Economic Crisis in East Asia : An Application of International Input-Output Analysis

This paper investigates the impacts of the 2008 economic crisis on industries in East Asia. By using the updated Asian international input-output table for 2008, the paper attempts to identify the transmission mechanism and the magnitude of impact of the crisis on industries in East Asia. The analyses reveal that the crisis significantly affected industrial output of the nine East Asian countries. In particular, the countries which are deeply involved in production networks were affected most seriously. Moreover, the analyses show that the impact was transmitted to East Asian industries considerably through the “triangular trade”, in which China imports parts and components from neighboring East Asian countries and then exports final products to the U.S. and EU markets.

Laser Shock Microformingof Thin Metal Sheets with ns Lasers

Continuous and long-pulse lasers have been used for the forming of metal sheets in macroscopic mechanical applications. However, for the manufacturing of micro-electromechanical systems (MEMS), the use of ns laser pulses provides a suitable parameter matching over an important range of sheet components that, preserving the short interaction time scale required for the predominantly mechanical (shock) induction of deformation residual stresses, allows for the successful processing of components in a medium range of miniaturization without appreciable thermal deformation.. In the present paper, the physics of laser shock microforming and the influence of the different experimental parameters on the net bending angle are presented.

Characterization Of Friction Stir Welded Joints Of Aa 2024-T351 - Native Vs. Treated By Laser Shock Processing

The paper presents a consistent set of results showing the ability of Laser Shock Processing (LSP) in modifying the overall properties of the Friction Stir Welded (FSW) joints made of AA 2024-T351. Based on laser beam intensities above 109 W/cm2 with pulse energies of several Joules and pulses durations of nanoseconds, LSP is able of inducing a compression residual stress field, improving the wear and fatigue resistance by slowing crack propagation and stress corrosion cracking, but also improving the overall behaviour of the structure. After the FSW and LSP procedures are briefly presented, the results of micro-hardness measurements and of transverse tensile tests, together with the corrosion resistance of the native joints vs. LSP treated are discussed. The ability of LSP to generate compressive residual stresses and to improve the behaviour of the FSW joints is underscored.

Direct-Drive Shock-Ignition for the Laser Megajoule.

We present a review of direct-drive shock ignition studies done as alternative for the Laser Mega-Joule to achieve high thermonuclear gain. One-dimensional analysis of HiPER-like Shock-ignited target designs is presented. It is shown that high gain can be achieved with shock ignition for designs which do not ignite only from the laser compression. Shock ignition is achieved for different targets of the fast ignition family which are driven by an absorbed energy between 100 kJ and 850kJ and deliver thermonuclear energies between 10-130 MJ. Shock-Ignition of Direct-Drive Double-Shell non-cryogenic target is also addressed. 2D results concerning the LMJ irradiation geometry are presented. Few systematic analyses are performed for the fuel assembly irradiation uniformity using the whole LMJ configuration or a part of the facility, and for the ignitor spike uniformity. Solutions for fuel assembly and shock ignition on LMJ using 2D calculations are presented. It is shown that high-gain shock-ignition is possible with intensity of each quad less than 1e15 W/cm2but low modes asymmetries displace the ignitor power in the spike towards higher powers.