Stress-induced Expression Research Articles

The DAXX tax: C9orf72 DNA repeat expansions drive gain- and loss-of-function pathology in c9FTD/ALS

In this issue of Neuron, Liu etal.1 identify DAXX as a C9orf72 hexanucleotide repeat expansion DNA-binding protein that initiates epigenetic modifications and chromatin remodeling, contributing to C9orf72 haploinsufficiency by inhibiting its stress-inducible expression and mediating both loss- and toxic gain-of-function pathology.

NF-κB Transcriptional Activity Indispensably Mediates Hypoxia–Reoxygenation Stress-Induced microRNA-210 Expression

Ischemia-reperfusion (I-R) injury is a cardinal pathophysiological hallmark of ischemic heart disease (IHD). Despite significant advances in the understanding of what causes I-R injury and hypoxia-reoxygenation (H-R) stress, viable molecular strategies that could be targeted for the treatment of the deleterious biochemical pathways activated during I-R remain elusive. The master hypoxamiR, microRNA-210 (miR-210), is a major determinant of protective cellular adaptation to hypoxia stress but exacerbates apoptotic cell death during cellular reoxygenation. While the hypoxia-induced transcriptional up-regulation of miR-210 is well delineated, the cellular mechanisms and molecular entities that regulate the transcriptional induction of miR-210 during the cellular reoxygenation phase have not been elucidated yet. Herein, in immortalized AC-16 cardiomyocytes, we delineated the indispensable role of the ubiquitously expressed transcription factor, NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) in H-R-induced miR-210 expression during cellular reoxygenation. Using dominant negative and dominant active expression vectors encoding kinases to competitively inhibit NF-κB activation, we elucidated NF-κB activation as a significant mediator of H-R-induced miR-210 expression. Ensuing molecular assays revealed a direct NF-κB-mediated transcriptional up-regulation of miR-210 expression in response to the H-R challenge that is characterized by the NF-κB-mediated reorchestration of the entire repertoire of histone modification changes that are a signatory of a permissive actively transcribed miR-210 promoter. Our study confers a novel insight identifying NF-κB as a potential novel molecular target to combat H-R-elicited miR-210 expression that fosters augmented cardiomyocyte cell death.

Traumatic Stress-Induced RNA Expression Signatures In Rats Reveal Insights Into Fkbp5-Mediated Posttraumatic Pain Development

Traumatic Stress-Induced RNA Expression Signatures In Rats Reveal Insights Into Fkbp5-Mediated Posttraumatic Pain Development

A PRECLINICAL MODEL OF STENOTIC CROHN’S DISEASE FOR THE STUDY OF MECHANISMS OF MOTILITY DYSFUNCTION IN IBD

Motility dysfunction is present in patients with stenotic Crohn’s disease (CD) and contributes to cramps, diarrhea and constipation. However, mechanisms for motility dysfunction in CD are incompletely understood. Mechanical stress, i.e. inflammatory infiltration, edema, stenosis, and distention, is commonly encountered in CD. We tested the hypothesis in a rat model of stenotic CD that mechanical stress-induced expression of mechano-sensitive genes such as cyclo-oxygenase-2 (COX-2) may account for motility dysfunction in the inflammation site and the mechanically distended site prior to inflammation.

The Role of Ketogenic Diet in the Management of Diabetes and Overcome it’s Eff ect: A Review

The ketogenic diet, which is low in carbohydrates, may help diabetic people reduce their medication and improve their glycemia. Obesity and diabetes are two of the most well-known metabolic diseases connected with unfortunate dietary patterns and a stationary way of life. In the direst outcome imaginable, metabolic issues are a contributing reason to a wide range of diseases. The ketogenic diet replaces glucose with ketone bodies and is useful in various conditions, including metabolic disorders, epileptic seizures, peripheral neuropathy, kidney cancer, and skeletal muscle atrophy. Obesity is connected to a higher risk of type two diabetes. In obese people with glucose intolerance, it has been discovered that successful weight management and dietary adjustments, particularly in terms of carbohydrate content and glycemic index, have favorable benefi ts. We’ve already demonstrated that a ketogenic diet could help you to lose weight. Furthermore, even in hyperlipidemic obese people, it improves cardiac risk factors. For a period of 56 weeks, the eff ect of low carbs or ketogenic diet was evaluated in patients having obesity with high glucose levels of blood to those with moderate blood glucose levels in this study. In many circumstances, excess insulin treatment and protein limitation slow the onset of diabetic nephropathy, and very few therapies are known to reverse nephropathy. The Keto diet regulates glucose and insulin levels in the body, making it an effi cient diabetes treatment. As a result, the keto diet can be used to demonstrate the gap between diabetes treatment and obesity. Obesity is connected to a higher risk diabetes of type 2. In obese people with glucose intolerance, it has been discovered that successful weight management and dietary adjustments, particularly in carbohydrate content and glycemic index, have favorable eff ects. Diabetic nephropathy was entirely restored after two months of following a ketogenic diet, as measured by albumin/creatinine ratios and stress-induced gene expression. On the other hand, histological evidence of nephropathy was only partially reversed. This shows that diabetic nephropathy can be restored with a relatively straightforward dietary change. It’s still unclear if the ketogenic diet’s benefi cial eff ects are mediated by lower glucose metabolism. Glycemic levels are aff ected by dietary changes. The low-carb ketogenic diet was found to be quite promising in managing diabetic mellitus in preliminary investigations.

Spatial Proteome Reorganization of a Photosynthetic Model Cyanobacterium in Response to Abiotic Stresses.

Spatial proteome reorganization in response to a changing environment represents a different layer of adaptation mechanism in addition to differential expression of a subset of stress responsive genes in photosynthetic organisms. Profiling such reorganization events is critically important to extend our understanding how photosynthetic organisms adapt to adverse environments. Thus, we treated a unicellular photosynthetic model cyanobacterium, Synechocystis sp. PCC 6803 (hereafter referred to as Synechocystis), with five different types of abiotic stresses including nitrogen starvation, iron deficiency, cold, heat, and darkness, and systematically identified proteins showing stress-induced differential expression and/or redistribution between the membrane and the soluble fractions using a quantitative proteomics approach. A number of proteins showing such a redistribution in response to a single or multiple types of abiotic stresses were identified. These include 12 ribosomal proteins displaying unanimous cold-induced redistribution to the membrane and the protein FurA, a master regulator of iron acquisition, displaying iron deficiency- and nitrogen starvation-induced redistribution to the membrane. Such findings shed light on a novel regulatory mechanism underlying the corresponding stress responses, and establish the results in the present study as an important resource for future studies intended to understand how photosynthetic organisms cope with adverse environments.

GLI1, a novel target of the ER stress regulator p97/VCP, promotes ATF6f-mediated activation of XBP1

Upon accumulation of improperly folded proteins in the Endoplasmic Reticulum (ER), the Unfolded Protein Response (UPR) is triggered to restore ER homeostasis. The induction of stress genes is a sine qua non condition for effective adaptive UPR. Although this requirement has been extensively described, the mechanisms underlying this process remain in part uncharacterized. Here, we show that p97/VCP, an AAA+ ATPase known to contribute to ER stress-induced gene expression, regulates the transcription factor GLI1, a primary effector of Hedgehog (Hh) signaling. Under basal (non-ER stress) conditions, GLI1 is repressed by a p97/VCP-HDAC1 complex while upon ER stress GLI1 is induced through a mechanism requiring both USF2 binding and increase histone acetylation at its promoter. Interestingly, the induction of GLI1 was independent of ligand-regulated Hh signaling. Further analysis showed that GLI1 cooperates with ATF6f to induce promoter activity and expression of XBP1, a key transcription factor driving UPR. Overall, our work demonstrates a novel role for GLI1 in the regulation of ER stress gene expression and defines the interplay between p97/VCP, HDAC1 and USF2 as essential players in this process.

Genome-Wide Identification of B3 DNA-Binding Superfamily Members (ABI, HIS, ARF, RVL, REM) and Their Involvement in Stress Responses and Development in Camelina sativa

The B3 DNA-binding superfamily is a group of gene families that contain a B3 domain in their proteins. Members of this superfamily are involved in responses to stresses including salt, drought, and cold stress. The B3 DNA-binding superfamily has not been fully studied in Camelina sativa. A total of eighty-seven members of this superfamily were identified in C. sativa. The identified genes were placed into five groups based on a phylogenetic analysis. All the proteins were predicted to be nuclear. The RAV and ARF gene family had the most conserved motifs, with nine out of ten motifs being preserved, while the REM gene family was discovered to have the fewest, with just one conserved motif being present. The RAV and REM gene families showed the least protein–protein interactions. The CsARF5 and CsARF7 genes showed the highest potent interaction score with multiple auxin-responsive proteins. A qPCR analysis was carried out on six genes that showed stress-induced expression changes. CsREM17, CsREM5, and CsRAVL5 were discovered to be considerably increased in response to drought stress, while CsARF10, CsARF4, and CsREM34 were found to be downregulated to a large extent. The B3 DNA-binding superfamily regulates abscisic acid signaling, which in turn influences plant growth and stress resistance.

DNA-initiated epigenetic cascades driven by C9orf72 hexanucleotide repeat

The C9orf72 hexanucleotide repeat expansion (HRE) is the most frequent genetic cause of the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here, we describe the pathogenic cascades that are initiated by the C9orf72 HRE DNA. The HRE DNA binds to its protein partner DAXX and promotes its liquid-liquid phase separation, which is capable of reorganizing genomic structures. An HRE-dependent nuclear accumulation of DAXX drives chromatin remodeling and epigenetic changes such as histone hypermethylation and hypoacetylation in patient cells. While regulating global gene expression, DAXX plays a key role in the suppression of basal and stress-inducible expression of C9orf72 via chromatin remodeling and epigenetic modifications of the promoter of the major C9orf72 transcript. Downregulation of DAXX or rebalancing the epigenetic modifications mitigates the stress-induced sensitivity of C9orf72-patient-derived motor neurons. These studies reveal a C9orf72 HRE DNA-dependent regulatory mechanism for both local and genomic architectural changes in the relevant diseases.

Multidimensional plasticity jointly contributes to rapid acclimation to environmental challenges during biological invasions.

Rapid plastic response to environmental changes, which involves extremely complex underlying mechanisms, is crucial for organismal survival during many ecological and evolutionary processes such as those in global change and biological invasions. Gene expression is among the most studied molecular plasticity, while co- or posttranscriptional mechanisms are still largely unexplored. Using a model invasive ascidian Ciona savignyi, we studied multidimensional short-term plasticity in response to hyper- and hyposalinity stresses, covering the physiological adjustment, gene expression, alternative splicing (AS), and alternative polyadenylation (APA) regulations. Our results demonstrated that rapid plastic response varied with environmental context, timescales, and molecular regulatory levels. Gene expression, AS, and APA regulations independently acted on different gene sets and corresponding biological functions, highlighting their nonredundant roles in rapid environmental adaptation. Stress-induced gene expression changes illustrated the use of a strategy of accumulating free amino acids under high salinity and losing/reducing them during low salinity to maintain the osmotic homoeostasis. Genes with more exons were inclined to use AS regulations, and isoform switches in functional genes such as SLC2a5 and Cyb5r3 resulted in enhanced transporting activities by up-regulating the isoforms with more transmembrane regions. The extensive 3'-untranslated region (3'UTR) shortening through APA was induced by both salinity stresses, and APA regulation predominated transcriptomic changes at some stages of stress response. The findings here provide evidence for complex plastic mechanisms to environmental changes, and thereby highlight the importance of systemically integrating different levels of regulatory mechanisms in studying initial plasticity in evolutionary trajectories.

Limited roles of Piezo mechanosensing channels in articular cartilage development and osteoarthritis progression

To investigate the role of Piezo1 and Piezo2 in surgically induced osteoarthritis (OA) in mice. Male conditional knockout (cKO) mice missing Piezo1 and Piezo2 in the joint using Gdf5-Cre transgenic mice were induced with post-traumatic OAby destabilization of the medial meniscus (DMM) of the right knee joint at 12 weeks of age. The severity of OA was histologically assessed at 24 weeks of age. OA-associated pain was evaluated by static weight bearing analysis. Additionally, articular chondrocytes isolated from cKO mice were exposed to fluid flow shear stress (FFSS) to evaluate the expression of OA-associated genes. Mice with conditional deletion of Piezo1 and Piezo2 showed normal joint development with no overt histological changes in the knee joint at 12 weeks and 24 weeks. DMM surgery induced moderate to severe OA in both control and cKO mice (median OARSI score: control, 4.67; cKO, 4.23, P=0.3082), although a few cKO mice showed milder OA. Pain assessment by static weight-bearing analysis suggested Piezo ablation in the joint has no beneficial effects on pain. FFSS increased the expression of OA-related genes both in control and cKO mice to similar extents. Piezo1 and Piezo2 are not essential for normal joint development. Genetic ablation of Piezo channels did not confer evident protective effects on OA progression in mice. In vitro data suggests that different mechanotransducers other than Piezo channels mediate FFSS in mechanical stress-induced gene expression.

Effects of footshock stress on social behavior and neuronal activation in the medial prefrontal cortex and amygdala of male and female mice.

Social behavior is complex and fundamental, and its deficits are common pathological features for several psychiatric disorders including anxiety, depression, and posttraumatic stress disorder. Acute stress may have a negative impact on social behavior, and these effects can vary based on sex. The aim of this study was to explore the effect of acute footshock stress, using analogous parameters to those commonly used in fear conditioning assays, on the sociability of male and female C57BL/6J mice in a standard social approach test. Animals were divided into two main groups of footshock stress (22 male, 24 female) and context exposed control (23 male and 22 female). Each group had mice that were treated intraperitoneally with either the benzodiazepine-alprazolam (control: 10 male, 10 female; stress: 11 male, 11 female), or vehicle (control: 13 male, 12 female; stress: 11 male, 13 female). In all groups, neuronal activation during social approach was assessed using immunohistochemistry against the immediate early gene product cFos. Although footshock stress did not significantly alter sociability or latency to approach a social stimulus, it did increase defensive tail-rattling behavior specifically in males (p = 0.0022). This stress-induced increase in tail-rattling was alleviated by alprazolam (p = 0.03), yet alprazolam had no effect on female tail-rattling behavior in the stress group. Alprazolam lowered cFos expression in the medial prefrontal cortex (p = 0.001 infralimbic area, p = 0.02 prelimbic area), and social approach induced sex-dependent differences in cFos activation in the ventromedial intercalated cell clusters (p = 0.04). Social approach following stress-induced cFos expression was positively correlated with latency to approach and negatively correlated with sociability in the prelimbic area and multiple amygdala subregions (all p < 0.05). Collectively, our results suggest that acute footshock stress induces sex-dependent alterations in defensiveness and differential patterns of cFos activation during social approach.

Expression Profile of miRNA from High, Middle, and Low Stress-Responding Sheep during Bacterial Endotoxin Challenge.

Animals respond to stress by activating a wide array of physiological and behavioral responses that are collectively referred to as the stress response. MicroRNAs (miRNAs) are small, noncoding RNAs that play key roles in the regulation of homeostasis. There are many reports demonstrating examples of stress-induced miRNA expression profiles. The aim of this study was to determine the circulatory miRNA profile of variable stress-responding lambs (n = 112) categorized based on their cortisol levels as high (HSR, 336.2 ± 27.9 nmol/L), middle (MSR, 147.3 ±9.5 nmol/L), and low (LSR, 32.1 ± 10.4 nmol/L) stress responders post-LPS challenge (400 ng/kg iv). Blood was collected from the jugular vein at 0 (T0) and 4 h (T4) post-LPS challenge, and miRNAs were isolated from four animals from each group. An array of 84 miRNAs and 6 individual miRNAs were evaluated using qPCR. Among 90 miRNAs, there were 48 differentially expressed (DE) miRNAs (log fold change (FC) > 2 < log FC) in the HSR group, 46 in the MSR group, and 49 in the LSR group compared with T0 (control) samples. In the HSR group, three miRNAs, miR-485-5p, miR-1193-5p, and miR-3957-5p were significantly (p < 0.05) upregulated, while seven miRNAs, miR-376b-3p, miR-376c-3p, miR-411b-5p, miR-376a-3p, miR-376b-3p, miR-376c-3p, and miR-381-3p, were downregulated (p < 0.05) as compared to the LSR and MSR groups. Functional analysis of DE miRNAs revealed their roles in Ras and MAPK signaling, cytokine signaling, the adaptive immune system, and transcription pathways in the HSR phenotype, implicating a hyper-induced acute-phase response. In contrast, in the LSR group, enriched pathways included glucagon signaling metabolic regulation, the transportation of amino acids and ions, and the integration of energy metabolism. Taken together, these results indicate variation in the acute-phase response to an immune stress challenge, and these miRNAs are implicated in regulating responses within cortisol-based phenotypes.

Derivatives of 3, 4, 5-Trimethoxycinnamic Acid Ameliorate Stress-Induced Anxiety in Mice and Rats.

Stress is an overwhelming problem associated with neuronal damage leading to anxiety and depression. The compound 3, 4, 5-trimethoxycinnamic acid (TMCA) has shown anti-stress effects; however, its derivatives remained unknown for their anxiolytic properties. Here, therefore, we investigated derivatives of TMCA (dTMCA) for their anxiolytic effects using immobilization and electric shock-induced stress in rats. Derivatives of TMCA ameliorated anxiety in mice and rats revealed by extended period of time spent in the open arms of elevated plus maze. Stress-mediated repression of tyrosine hydroxylase (TH) protein expression in the amygdala regions of rat brain and dopamine levels in the PC12 cells was restored by two selected derivatives (TMCA#5 and TMCA#9). Unlike TH expression, stress-induced protein expression of phospho-extracellular signal-regulated kinase (pERK) was unaffected by both derivatives in rats. Given the preferential inhibitory activity of dTMCA on dopamine and serotonin receptors, serotonergic road map of cellular signaling could be their target for anxiolytic effects. Thus, dTMCA would be promising agents to prevent neuronal damage associated with rampant stressful conditions.

A PRECLINICAL MODEL OF STENOTIC CROHN’S DISEASE FOR THE STUDY OF MECHANISMS OF MOTILITY DYSFUNCTION IN IBD

Abstract BACKGROUND AND AIMS Motility dysfunction is present in patients with stenotic Crohn’s disease (CD) and contributes to cramps, diarrhea and constipation. However, mechanisms for motility dysfunction in CD are incompletely understood. Mechanical stress, i.e. inflammatory infiltration, edema, stenosis, and distention, is commonly encountered in CD. We tested the hypothesis in a rat model of stenotic CD that mechanical stress-induced expression of mechano-sensitive genes such as cyclo-oxygenase-2 (COX-2) may account for motility dysfunction in the inflammation site and the mechanically distended site prior to inflammation. ANIMAL MODEL AND METHODS Rat model of CD-like colitis was induced by intracolonic instillation of TNBS 65 mg/kg in 250 µL of 40% ethanol. Control rats were treated with intracolonic saline instillation. When rats were fed with regular pellet food, instillation of TNBS led to localized transmural inflammation with stenosis in the instillation site (site I) and lumen distention in the colon segment proximal to inflammation (site P). However, when rats were fed exclusively with liquid diet, inflammation-associated stenosis and mechanical distention was eliminated. Rats were euthanized by 7 days. RESULTS 1) Compared to controls, COX-2 mRNA expression was increased by 13.1(± 2.5)-fold in site I and 16.6(±4.3)-fold in the mechanically distended site P, but not in the non-distended site D. This data indicates a mechanosensitive mechanism in COX-2 expression. If mechanical distention in CD was prevented by feeding rats exclusively with clear liquid diet, the increase of COX-2 mRNA expression was attenuated in sites I and P. Further investigation showed that increased expression of COX-2 occurs mainly in the colonic smooth muscle cells (SMC) of the distended colon. 2) We found that COX-2 expression was robustly induced by mechanical stretch (18% static stretch) in vitro in rat colonic smooth muscle cells (SMC). 3) Inflammation led to significant suppression of colon circular muscle contractility not only in site I but also in distended site P of CD-like rats. Muscle contractility in site D was not significantly affected by inflammation. Moreover, clear liquid diet treatment significantly improved muscle contractility in sites P and I. 4) To determine the role of COX-2 in smooth muscle dysfunction in the CD model, COX-2 inhibitor NS-398 (5 mg/kg, i.p. daily) was administered in control and CD rats. We found that NS-398 significantly improved muscle contractility in CD rats in sites P and I. CONCLUSIONS Our study found that up-regulation of COX-2 expression in stenotic CD-like colitis is dependent on mechanical stress. COX-2 inhibitor improved colon smooth muscle contractility in colitis. Thus, mechanosensitive expression of COX-2 may play a critical role in smooth muscle dysfunction and dysmotility in CD.

LSD1 defines the fiber type-selective responsiveness to environmental stress in skeletal muscle.

Skeletal muscle exhibits remarkable plasticity in response to environmental cues, with stress-dependent effects on the fast-twitch and slow-twitch fibers. Although stress-induced gene expression underlies environmental adaptation, it is unclear how transcriptional and epigenetic factors regulate fiber type-specific responses in the muscle. Here, we show that flavin-dependent lysine-specific demethylase-1 (LSD1) differentially controls responses to glucocorticoid and exercise in postnatal skeletal muscle. Using skeletal muscle-specific LSD1-knockout mice and in vitro approaches, we found that LSD1 loss exacerbated glucocorticoid-induced atrophy in the fast fiber-dominant muscles, with reduced nuclear retention of Foxk1, an anti-autophagic transcription factor. Furthermore, LSD1 depletion enhanced endurance exercise-induced hypertrophy in the slow fiber-dominant muscles, by induced expression of ERRγ, a transcription factor that promotes oxidative metabolism genes. Thus, LSD1 serves as an 'epigenetic barrier' that optimizes fiber type-specific responses and muscle mass under the stress conditions. Our results uncover that LSD1 modulators provide emerging therapeutic and preventive strategies against stress-induced myopathies such as sarcopenia, cachexia, and disuse atrophy.

Cloning and characterization of vacuolar Na+/H+ antiporter gene promoter from wheat (Triticum aestivum) cv. Kharchia Local

Soil salinity is one of the significant abiotic stresses in plants. Few efficient stress-inducible promoters from plants, especially from monocotyledonous crops like wheat (Triticum aestivum L.) are available to drive stress-inducible expression of transgene. Therefore, an experiment was conducted at ICAR-National Institute for Plant Biotechnology, New Delhi, during 2017–18 to clone and characterize stress-inducible promoter from wheat. Results found a 418 bp long NHX1 promoter (TaUNHX) from a bread wheat landrace, Kharchia Local. In silico analysis of TaUNHX predicted several cis-acting regulatory elements, including CAAT, DOF, GATA motifs and some essential stress-responsive elements. To analyze the activity of TaUNHX, Agrobacterium-mediated transient GUS assay in tobacco (Nicotiana tabacum L.) leaves and stems was studied under various abiotic stresses like salt (300 mM NaCl), drought (20% PEG) and ABA (100 μM). Stress-responsive nature of this promoter can be used to drive the expression of transgene following exposure to various stresses. The study's finding has significant implications as the characterized promoter can be used to develop transgenics where the transgene expression can be induced following exposure to stress.

Overexpression of a DUF740 family gene (LOC_Os04g59420) imparts enhanced climate resilience through multiple stress tolerance in rice.

Functional characterization of stress-responsive genes through the analysis of transgenic plants is a standard approach to comprehend their role in climate resilience and subsequently exploit them for sustainable crop improvement. In this study, we investigated the function of LOC_Os04g59420, a gene of DUF740 family (OsSRDP-Oryza sativa Stress Responsive DUF740 Protein) from rice, which showed upregulation in response to abiotic stress in the available global expression data, but is yet to be functionally characterized. Transgenic plants of the rice OsSRDP gene, driven by a stress-inducible promoter AtRd29A, were developed in the background of cv. Pusa Sugandh 2 (PS2) and their transgene integration and copy number were confirmed by molecular analysis. The three independent homozygous transgenic plants (AtRd29A::OsSRDP rice transformants) showed better resilience to drought, salinity, and cold stresses, but not heat stress, as compared to the non-transformed PS2, which corresponded with their respective relative transcript abundance for OsSRDP. Transgenic plants maintained higher RWC, photosynthetic pigments, and proline accumulation under drought and salinity stresses. Furthermore, they exhibited less accumulation of reactive oxygen species (ROS) than PS2 under drought stress, as seen from the transcript abundance studies of the ROS genes. Under cold stress, OsSRDP transgenic lines illustrated minimal cell membrane injury compared to PS2. Additionally, the transgenic plants showed resistance to a virulent strain of rice blast fungus, Magnaporthe oryzae (M. oryzae). The promoter analysis of the gene in N22 and PS2 revealed the presence of multiple abiotic and biotic stress-specific motif elements supporting our observation on multiple stress tolerance. Based on bioinformatics studies, we identified four potential candidate interaction partners for LOC_Os04g59420, of which two genes (LOC_Os05g09640 and LOC_Os06g50370) showed co-expression under biotic and drought stress along with OsSRDP. Altogether, our findings established that stress-inducible expression of OsSRDP can significantly enhance tolerance to multiple abiotic stresses and a biotic stress.

Single-cell RNA sequencing reveals the suppressive effect of PPP1R15A inhibitor Sephin1 in antitumor immunity

Protein phosphatase 1 regulatory subunit 15A (PPP1R15A) is an important factor in the integrated stress response (ISR) in mammals and may play a crucial role in tumorigenesis. In our studies, we found an inhibitor of PPP1R15A, Sephin1, plays a protumorigenic role in mouse tumor models. By analyzing the single-cell transcriptome data of the mouse tumor models, we found that in C57BL/6 mice, Sephin1 treatment could lead to higher levels of ISR activity and lower levels of antitumor immune activities. Specifically, Sephin1 treatment caused reductions in antitumor immune cell types and lower expression levels of cytotoxicity-related genes. In addition, Tcell receptor (TCR) repertoire analysis demonstrated that the clonal expansion of tumor-specific Tcells was inhibited by Sephin1. A special TCR+ macrophage subtype in tumor was identified to be significantly depleted upon Sephin1 treatment, implying its key antitumor role. These results suggest that PPP1R15A has the potential to be an effective target for tumor therapy.

Comparative transcriptomic analysis of the gene expression and underlying molecular mechanism of submergence stress response in orchardgrass roots.

Submergence stress creates a hypoxic environment. Roots are the first plant organ to face these low-oxygen conditions, which causes damage and affects the plant growth and yield. Orchardgrass (Dactylis glomerata L.) is one of the most important cold-season forage grasses globally. However, their submergence stress-induced gene expression and the underlying molecular mechanisms of orchardgrass roots are still unknown. Using the submergence-tolerant 'Dianbei' and submergence-sensitive 'Anba', the transcriptomic analysis of orchardgrass roots at different time points of submergence stress (0 h, 8 h, and 24 h) was performed. We obtained 118.82Gb clean data by RNA-Seq. As compared with the control, a total of 6663 and 9857 differentially expressed genes (DEGs) were detected in Dianbei, while 7894 and 11215 DEGs were detected in Anba at 8 h and 24 h post-submergence-stress, respectively. Gene Ontology (GO) enrichment analysis obtained 986 terms, while Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis obtained 123 pathways. Among them, the DEGs in plant hormones, mitogen-activated protein kinase (MAPK) and Ca2+ signal transduction were significantly differentially expressed in Dianbei, but not in Anba. This study was the first to molecularly elucidate the submergence stress tolerance in the roots of two orchardgrass cultivars. These findings not only enhanced our understanding of the orchardgrass submergence tolerance, but also provided a theoretical basis 36 for the cultivation of submergence-tolerant forage varieties.