scholarly journals The amino acid substitution affects cellular response to mistranslation

2021 ◽  
Author(s):  
Matthew D. Berg ◽  
Yanrui Zhu ◽  
Bianca Y. Ruiz ◽  
Raphaël Loll-Krippleber ◽  
Joshua Isaacson ◽  
...  
Keyword(s):  
Amino Acid ◽  
Heat Shock ◽  
Amino Acid Substitution ◽  
Heat Shock Response ◽  
Cellular Response ◽  
Genetic Interactions ◽  
Similar Frequency ◽  
Proteotoxic Stress ◽  
Shock Response ◽  
The Impact

Mistranslation, the mis-incorporation of an amino acid not specified by the standard genetic code, occurs in all organisms. tRNA variants that increase mistranslation arise spontaneously and engineered tRNAs can achieve mistranslation frequencies approaching 10% in yeast and bacteria. Interestingly, human genomes contain tRNA variants with the potential to mistranslate. Cells cope with increased mistranslation through multiple mechanisms, though high levels cause proteotoxic stress. The goal of this study was to compare the genetic interactions and the impact on transcriptome and cellular growth of two tRNA variants that mistranslate at a similar frequency but create different amino acid substitutions in Saccharomyces cerevisiae. One tRNA variant inserts alanine at proline codons whereas the other inserts serine for arginine. Both tRNAs decreased growth rate, with the effect being greater for arginine to serine than for proline to alanine. The tRNA that substituted serine for arginine resulted in a heat shock response. In contrast, heat shock response was minimal for proline to alanine substitution. Further demonstrating the significance of the amino acid substitution, transcriptome analysis identified unique up- and downregulated genes in response to each mistranslating tRNA. Number and extent of negative synthetic genetic interactions also differed depending upon type of mistranslation. Based on the unique responses observed for these mistranslating tRNAs, we predict that the potential of mistranslation to exacerbate diseases caused by proteotoxic stress depends on the tRNA variant. Furthermore, based on their unique transcriptomes and genetic interactions, different naturally occurring mistranslating tRNAs have the potential to negatively influence specific diseases.

scholarly journals The amino acid substitution affects cellular response to mistranslation

2021 ◽  
Author(s):  
Matthew D Berg ◽  
Yanrui Zhu ◽  
Bianca Y Ruiz ◽  
Raphaël Loll-Krippleber ◽  
Joshua Isaacson ◽  
...  
Keyword(s):  
Amino Acid ◽  
Heat Shock ◽  
Amino Acid Substitution ◽  
Heat Shock Response ◽  
Cellular Response ◽  
Genetic Interactions ◽  
Similar Frequency ◽  
Proteotoxic Stress ◽  
Shock Response ◽  
The Impact

Abstract Mistranslation, the mis-incorporation of an amino acid not specified by the “standard” genetic code, occurs in all organisms. tRNA variants that increase mistranslation arise spontaneously and engineered tRNAs can achieve mistranslation frequencies approaching 10% in yeast and bacteria. Interestingly, human genomes contain tRNA variants with the potential to mistranslate. Cells cope with increased mistranslation through multiple mechanisms, though high levels cause proteotoxic stress. The goal of this study was to compare the genetic interactions and the impact on transcriptome and cellular growth of two tRNA variants that mistranslate at a similar frequency but create different amino acid substitutions in Saccharomyces cerevisiae. One tRNA variant inserts alanine at proline codons whereas the other inserts serine for arginine. Both tRNAs decreased growth rate, with the effect being greater for arginine to serine than for proline to alanine. The tRNA that substituted serine for arginine resulted in a heat shock response. In contrast, heat shock response was minimal for proline to alanine substitution. Further demonstrating the significance of the amino acid substitution, transcriptome analysis identified unique up- and downregulated genes in response to each mistranslating tRNA. Number and extent of negative synthetic genetic interactions also differed depending upon type of mistranslation. Based on the unique responses observed for these mistranslating tRNAs, we predict that the potential of mistranslation to exacerbate diseases caused by proteotoxic stress depends on the tRNA variant. Furthermore, based on their unique transcriptomes and genetic interactions, different naturally occurring mistranslating tRNAs have the potential to negatively influence specific diseases.

scholarly journals Efficient Heat Shock Response Affects Hyperthermia-Induced Radiosensitization in a Tumor Spheroid Control Probability Assay

Cancers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 3168
Author(s):  
Oleg Chen ◽  
Soňa Michlíková ◽  
Lisa Eckhardt ◽  
Marit Wondrak ◽  
Adriana M. De Mendoza ◽  
...  
Keyword(s):  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Cellular Stress Response ◽  
Proteotoxic Stress ◽  
Shock Response ◽  
Pre Treatment ◽  
Shock Proteins ◽  
The Impact

Hyperthermia (HT) combined with irradiation is a well-known concept to improve the curative potential of radiotherapy. Technological progress has opened new avenues for thermoradiotherapy, even for recurrent head and neck squamous cell carcinomas (HNSCC). Preclinical evaluation of the curative radiosensitizing potential of various HT regimens remains ethically, economically, and technically challenging. One key objective of our study was to refine an advanced 3-D assay setup for HT + RT research and treatment testing. For the first time, HT-induced radiosensitization was systematically examined in two differently radioresponsive HNSCC spheroid models using the unique in vitro “curative” analytical endpoint of spheroid control probability. We further investigated the cellular stress response mechanisms underlying the HT-related radiosensitization process with the aim to unravel the impact of HT-induced proteotoxic stress on the overall radioresponse. HT disrupted the proteome’s thermal stability, causing severe proteotoxic stress. It strongly enhanced radiation efficacy and affected paramount survival and stress response signaling networks. Transcriptomics, q-PCR, and western blotting data revealed that HT + RT co-treatment critically triggers the heat shock response (HSR). Pre-treatment with chemical chaperones intensified the radiosensitizing effect, thereby suppressing HT-induced Hsp27 expression. Our data suggest that HT-induced radiosensitization is adversely affected by the proteotoxic stress response. Hence, we propose the inhibition of particular heat shock proteins as a targeting strategy to improve the outcome of combinatorial HT + RT.

Development of the embryonic heat shock response and the impact of repeated thermal stress in early stage lake whitefish ( Coregonus clupeaformis ) embryos

2017 ◽  
Vol 69 ◽  
pp. 294-301 ◽  
Author(s):  
Lindy M. Whitehouse ◽  
Chance S. McDougall ◽  
Daniel I. Stefanovic ◽  
Douglas R. Boreham ◽  
Christopher M. Somers ◽  
...  
Keyword(s):  
Thermal Stress ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Early Stage ◽  
Coregonus Clupeaformis ◽  
Lake Whitefish ◽  
Shock Response ◽  
The Impact

scholarly journals The Quinone Methide Aurin Is a Heat Shock Response Inducer That Causes Proteotoxic Stress and Noxa-dependent Apoptosis in Malignant Melanoma Cells

2014 ◽  
Vol 290 (3) ◽  
pp. 1623-1638 ◽  
Author(s):  
Angela L. Davis ◽  
Shuxi Qiao ◽  
Jessica L. Lesson ◽  
Montserrat Rojo de la Vega ◽  
Sophia L. Park ◽  
...  
Keyword(s):  
Heat Shock ◽  
Malignant Melanoma ◽  
Heat Shock Response ◽  
Melanoma Cells ◽  
Quinone Methide ◽  
Proteotoxic Stress ◽  
Shock Response

scholarly journals Chemical-Genetic Interactions with the Proline Analog L-Azetidine-2-Carboxylic Acid in Saccharomyces cerevisiae

2020 ◽  
Vol 10 (12) ◽  
pp. 4335-4345
Author(s):  
Matthew D. Berg ◽  
Yanrui Zhu ◽  
Joshua Isaacson ◽  
Julie Genereaux ◽  
Raphaël Loll-Krippleber ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Carboxylic Acid ◽  
Cellular Response ◽  
Protein Quality ◽  
Genetic Interaction ◽  
Genetic Interactions ◽  
Temperature Sensitive ◽  
Chemical Genetic ◽  
Proteotoxic Stress

Non-proteinogenic amino acids, such as the proline analog L-azetidine-2-carboxylic acid (AZC), are detrimental to cells because they are mis-incorporated into proteins and lead to proteotoxic stress. Our goal was to identify genes that show chemical-genetic interactions with AZC in Saccharomyces cerevisiae and thus also potentially define the pathways cells use to cope with amino acid mis-incorporation. Screening the yeast deletion and temperature sensitive collections, we found 72 alleles with negative chemical-genetic interactions with AZC treatment and 12 alleles that suppress AZC toxicity. Many of the genes with negative chemical-genetic interactions are involved in protein quality control pathways through the proteasome. Genes involved in actin cytoskeleton organization and endocytosis also had negative chemical-genetic interactions with AZC. Related to this, the number of actin patches per cell increases upon AZC treatment. Many of the same cellular processes were identified to have interactions with proteotoxic stress caused by two other amino acid analogs, canavanine and thialysine, or a mistranslating tRNA variant that mis-incorporates serine at proline codons. Alleles that suppressed AZC-induced toxicity functioned through the amino acid sensing TOR pathway or controlled amino acid permeases required for AZC uptake. Further suggesting the potential of genetic changes to influence the cellular response to proteotoxic stress, overexpressing many of the genes that had a negative chemical-genetic interaction with AZC suppressed AZC toxicity.

scholarly journals Depending on the stress, histone deacetylase inhibitors act as heat shock protein co-inducers in motor neurons and potentiate arimoclomol, exerting neuroprotection through multiple mechanisms in ALS models

2020 ◽  
Vol 25 (1) ◽  
pp. 173-191 ◽  
Author(s):  
Rachel Kuta ◽  
Nancy Larochelle ◽  
Mario Fernandez ◽  
Arun Pal ◽  
Sandra Minotti ◽  
...  
Keyword(s):  
Heat Shock ◽  
Histone Deacetylase ◽  
Motor Neurons ◽  
Heat Shock Response ◽  
Hdac Inhibitors ◽  
Dependent Manner ◽  
Hdac Inhibition ◽  
Proteotoxic Stress ◽  
Shock Response ◽  
Stress Dependent

AbstractUpregulation of heat shock proteins (HSPs) is an approach to treatment of neurodegenerative disorders with impaired proteostasis. Many neurons, including motor neurons affected in amyotrophic lateral sclerosis (ALS), are relatively resistant to stress-induced upregulation of HSPs. This study demonstrated that histone deacetylase (HDAC) inhibitors enable the heat shock response in cultured spinal motor neurons, in a stress-dependent manner, and can improve the efficacy of HSP-inducing drugs in murine spinal cord cultures subjected to thermal or proteotoxic stress. The effect of particular HDAC inhibitors differed with the stress paradigm. The HDAC6 (class IIb) inhibitor, tubastatin A, acted as a co-inducer of Hsp70 (HSPA1A) expression with heat shock, but not with proteotoxic stress induced by expression of mutant SOD1 linked to familial ALS. Certain HDAC class I inhibitors (the pan inhibitor, SAHA, or the HDAC1/3 inhibitor, RGFP109) were HSP co-inducers comparable to the hydroxyamine arimoclomol in response to proteotoxic stress, but not thermal stress. Regardless, stress-induced Hsp70 expression could be enhanced by combining an HDAC inhibitor with either arimoclomol or with an HSP90 inhibitor that constitutively induced HSPs. HDAC inhibition failed to induce Hsp70 in motor neurons expressing ALS-linked mutant FUS, in which the heat shock response was suppressed; yet SAHA, RGFP109, and arimoclomol did reduce loss of nuclear FUS, a disease hallmark, and HDAC inhibition rescued the DNA repair response in iPSC-derived motor neurons carrying the FUSP525Lmutation, pointing to multiple mechanisms of neuroprotection by both HDAC inhibiting drugs and arimoclomol.

scholarly journals Systemic activation coordinates the heat shock response of the insulin/IGF-1 pathway in Caenorhabditis elegans

2017 ◽  
Author(s):  
Ronen B Kopito ◽  
Kathie Watkins ◽  
Erel Levine
Keyword(s):  
Caenorhabditis Elegans ◽  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Cellular Response ◽  
Precise Control ◽  
Cellular Processes ◽  
Shock Response ◽  
Stressed Cells ◽  
Shock Proteins

Exposure to high temperatures has an adverse effect on cellular processes and results in activation of the cellular heat shock response (HSR), a highly conserved program of inducible genes to maintain protein homeostasis1. The insulin/IGF-1 signaling (IIS) pathway, which has diverse roles from metabolism to stress response and longevity, is activated as part of the HSR2–4. Recent evidence suggest that the IIS pathway is able to affect proteostasis non-autonomously5,6, yet it is not known if it is activated autonomously in stressed cells or systemically as part of an organismic program. In Caenorhabditis elegans, the single forkhead box O (FOXO) homologue DAF-16 functions as the major target of the IIS pathway7 and, together with the heat-shock factor HSF-1, induce the expression of small heat shock proteins in response to heat shock8–10,3. Here we use a novel microfluidic device that allows precise control of the spatiotemporal temperature profile to show that cellular activation of DAF-16 integrates local temperature sensation with systemic signals. We demonstrate that DAF-16 activation in head sensory neurons is essential for DAF-16 activation in other tissues, but show that no known thermosensory neuron is individually required. Our findings demonstrate that systemic and cell-autonomous aspects of stress response act together to facilitate a coordinated cellular response at the organismic level.

scholarly journals Heat shock response pathways regulate stimulus-specificity and sensitivity of NF-κB signalling to temperature stress

2019 ◽  
Author(s):  
Anna Paszek ◽  
Małgorzata Kardyńska ◽  
James Bagnall ◽  
Jarosław Śmieja ◽  
David G. Spiller ◽  
...  
Keyword(s):  
Heat Shock ◽  
Cross Talk ◽  
Heat Shock Response ◽  
Cellular Response ◽  
Breast Adenocarcinoma ◽  
Heat Shock Factor 1 ◽  
Induced Responses ◽  
Specificity And Sensitivity ◽  
Shock Response ◽  
Stimulus Specificity

AbstractAbility to adapt to temperature changes trough the Heat Shock Response (HSR) pathways is one of the most fundamental and clinically relevant cellular response systems. Here we report that Heat Shock (HS) induces a temporally-coordinated and stimulus-specific adaptation of the signalling and gene expression responses of the Nuclear Factor κB (NF-κB) transcription factor. We show that exposure of MCF7 breast adenocarcinoma cells to 43°C 1h HS inhibits the immediate signalling response to pro-inflammatory Interleukin 1β (IL1β) and Tumour Necrosis Factor α (TNFα) cytokines. Within 4h after HS treatment IL1β-induced responses return to normal levels, but the recovery of the TNFα-induced responses is delayed. Using siRNA knock-down of Heat Shock Factor 1 and mathematical modelling we show that the stimulus-specificity is conferred via the Inhibitory κB kinase signalosome, with HSR differentially controlling individual cytokine transduction pathways. Finally, using a novel mathematical model we predict and experimentally validate that the HSR cross-talk confers differential cytokine sensitivity of the NF-κB system to a range of physiological and clinically-relevant temperatures. This quantitative understanding of NF-κB and HSR cross-talk mechanisms is fundamentally important for the potential improvement of current hyperthermia protocols.

scholarly journals Proteotoxic stress of cancer: Implication of the heat-shock response in oncogenesis

2012 ◽  
Vol 227 (8) ◽  
pp. 2982-2987 ◽  
Author(s):  
Chengkai Dai ◽  
Siyuan Dai ◽  
Junyue Cao
Keyword(s):  
Heat Shock ◽  
Heat Shock Response ◽  
Proteotoxic Stress ◽  
Shock Response

scholarly journals Therapeutic Targeting of HSP90 in AML and ALL

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4680-4680
Author(s):  
Sanil Bhatia ◽  
Heinz Ahlert ◽  
Benedikt Frieg ◽  
Arndt Borkhardt ◽  
Holger Gohlke ◽  
...  
Keyword(s):  
Heat Shock ◽  
Small Molecule ◽  
Heat Shock Response ◽  
Cellular Response ◽  
Small Molecule Inhibitor ◽  
Protective Mechanism ◽  
Hsp90 Inhibitors ◽  
C Terminus ◽  
Molecule Inhibitor ◽  
Shock Response

Abstract Background: Even with the prevalent usage of specified treatment protocols, treatment gap remains and besides that the conventional therapies used routinely inflict significant toxicity due to low specificity. Therefore, the development of novel targeted therapies which are active against resistant leukemia subtypes and at the same time offer low toxicity in patients is of high importance. We, therefore, aimed to develop and characterize novel precision compounds, which target oncogene stabilization via HSP90 axis. Aims: We have previously developed a novel peptidomimetic HSP90 inhibitor (AX) which was active as a pan-leukemia inhibitor against LSCs without inducing any Heat Shock Resoponse (HSR). However due to peptidomimetic nature and high molecular weight, the clinical implication of AX was limited. Therefore, using the previous knowledge we focused on developing second generation of small molecule inhibitor against the C-terminal dimerization of HSP90 with better efficacy and clinical potential. Methods: We have generated the library of small molecule inhibitor targeting HSP90 C-terminus and selected the most potent analogue (VWK147) depending upon its higher potency against leukemic/cancerous cells. The specificity of VWK147 was further evaluated by microscale thermophoresis (MST), cell-based luciferase refolding assay, 2D NMR spectroscopy, analytical ultracentrifugation and molecular dynamics simulations. Results: HSP90 act as molecular chaperone and is highly expressed in several therapy-resistant leukemia subtypes thereby ensuring correct protein folding of several oncogenic proteins such as BCR-ABL1, FLT3-ITD and AKT. Therefore, targeting HSP90 could be a promising option in the treatment of therapy-refractory leukemia. Majority of available HSP90 inhibitors target the N-terminal domain thereby induce a protective mechanism called heat shock response (HSR), which potentially weakens the cytotoxic effect of HSP90 inhibitors and induce toxicity. We have now developed first in class small molecule HSP90 C-terminal dimerization inhibitor 'VWK147' through structure-based molecular design and chemical synthesis which specifically targets C-terminal dimerization of HSP90. Like AX, VWK147 destabilizes BCR-ABL oncoprotein and its related pro-oncogenic cellular response (involving proliferation, apoptosis and differentiation), effective in preclinical AML and TKI (2nd and 3rd generation) resistant cell line models in vitro and induces apoptosis in primary AML and BCR-ABL1+ BCP-ALL patient derived leukemic cell, without inducing any HSR. The next step would be to to evaluate its in vivo activity and pharmacodynamic profiling. Conclusion: Taken together, VWK147 represents a promising next step for future efforts towards the development of novel targeted HSP90 inhibitors to overcome drug resistance and reduce toxicity, especially for the treatment of relapsed/refractory ALL. References:Bhatia S, Diedrich D, Frieg B, et al. Targeting HSP90 dimerization via the C terminus is effective in imatinib-resistant CML and lacks the heat shock response. Blood. 2018;132(3):307-320.John C. Byrd. HSP90 inhibition without heat shock response. Blood commentary 2018. doi: https://doi.org/10.1182/blood-2018-05-850271. Disclosures No relevant conflicts of interest to declare.

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