Translational Signals Research Articles

EEF1A2 exacerbated insulin resistance in male skeletal muscle via PKCβ and ER stress.

Recent studies raise the possibility that eukaryotic translation elongation factor 1 alpha (eEF1A) may play a role in metabolism. One isoform, eEF1A2, is specifically expressed in skeletal muscle, heart and brain. It regulates translation elongation and signal transduction. Nonetheless, eEF1A2's function in skeletal muscle glucose metabolism remains unclear. In the present study, suppression subtractive hybridisation showed a decrease in Eef1a2 transcripts in the skeletal muscle of diabetic Mongolian gerbils. This was confirmed at mRNA and protein levels in hyperglycaemic gerbils, and in db/db and high-fat diet-fed mice. Further, this downregulation was independent of Eef1a2 promoter methylation. Interestingly, adeno-associated virus-mediated eEF1A2 overexpression in skeletal muscle aggravated fasting hyperglycaemia, hyperinsulinaemia and glucose intolerance in male diabetic gerbils but not in female gerbil models. The overexpression of eEF1A2 in skeletal muscle also resulted in promoted serum glucose levels and insulin resistance in male db/db mice. Up- and downregulation of eEF1A2 by lentiviral vector transfection confirmed its inhibitory effect on insulin-stimulated glucose uptake and signalling transduction in C2C12 myotubes with palmitate (PA)-induced insulin resistance. Furthermore, eEF1A2 bound PKCβ and increased its activation in the cytoplasm, whereas suppression of PKCβ by an inhibitor attenuated eEF1A2-mediated impairment of insulin sensitivity in insulin-resistant myotubes. Endoplasmic reticulum (ER) stress was elevated by eEF1A2, whereas suppression of ER stress or JNK partially restored insulin sensitivity in PA-treated myotubes. Additionally, eEF1A2 inhibited lipogenesis and lipid utilisation in insulin-resistant skeletal muscle. Collectively, we demonstrated that eEF1A2 exacerbates insulin resistance in male murine skeletal muscle via PKCβ and ER stress.

Nonstationary Heart Rate Variability during the Head-Down Tilt Test

Abstract—A new method has been proposed for processing the nonstationary rhythmogram signal recorded in the head-down tilt test (HDT). The method is based on the continuous wavelet transform of the heart rate signal with a frequency modulation component. The rhythmogram signal was shown to be a system of heart rhythm bursts of different frequency ranges. Quantitative indices were developed to analyze the system of bursts. The translational signal transfer method was used to eliminate the boundary effects that arise in analyses of the low-frequency components of the rhythmogram signal.

Developmental Cytoplasmic-to-Nuclear Translocation of RNA-Binding Protein HuR Is Required for Adult Neurogenesis

Although adult neurogenesis recapitulates processes that occur during embryonic development, it exhibits distinct characteristics from the embryonic counterpart. However, the intrinsic mechanism underlying the differential regulation of neurogenesis between these two stages remains unclear. Herein, we show that the ablation of RNA-binding protein HuR in NSCs impairs adult but not embryonic neurogenesis. HuR is predominantly expressed in the cytoplasm of embryonic NSCs but translocates into the nucleus of adult NSCs. Transcriptomic analysis of HuR-deficient adult NSCs revealed that HuR primarily regulates alternative splicing of numerous premRNA transcripts, including focal adhesion kinase (FAK). HuR-deficient adult NSCs generate increased FAK mRNA isoforms with shorter 5'-UTRs, leading to enhanced FAK mRNA translation and hyperactivated FAK signaling, and inhibition of FAK ameliorates defective adult neurogenesis and impaired hippocampus-dependent learning in HuR-deficient mice. These findings provide mechanistic insights into the differential regulation of embryonic and adult neurogenesis through developmental cytoplasmic-to-nuclear translocation of HuR.

Genomic survey of RNA recognition motif (RRM) containing RNA binding proteins from barley (Hordeum vulgare ssp. vulgare)

One of the major mechanisms of post-transcriptional gene regulation is achieved by proteins bearing well-defined sequence motifs involved in ‘RNA binding’. In eukaryotes, RNA binding proteins (RBPs) are key players of RNA metabolism that includes synthesis, processing, editing, modifying, transport, storage and stability of RNA. In plants, the family of RBPs is vastly expanded compared to other eukaryotes including humans. In this study we identified 363 RBPs in the barley genome. Gene ontology enrichment analysis of barley RBPs indicated these proteins were in all the major cellular compartments and associated with key biological processes including translation, splicing, seed development and stress signaling. Members with the classical RNA binding motifs such as the RNA recognition motif (RRM), KH domain, Helicase, CRM, dsRNA and Pumilio were identified in the repertoire of barley RBPs. Similar to Arabidopsis, the RRM containing RBPs were the most abundant in barley genome. In-depth analysis of the RRM containing proteins - polyA binding proteins, Ser/Arg rich proteins and Glycine-rich RBPs were undertaken. Reanalysis of the proteome dataset of various stages during barley malting identified 38 RBPs suggesting an important role for these proteins during the malting process. This survey provides a systematic analysis of barley RBPs and serves as the basis for the further functional characterization of this important family of proteins.

Cross-talk between ribosome biogenesis, translation, and mTOR in CD133+ 4/CD44+ prostate cancer stem cells.

To investigate the gene expression profile of CSCs and to explore the key pathways and specific molecular signatures involved in the characteristic of CSCs. CD133+ /CD44+ CSCs and bulk population (non-CSCs) were isolated from DU-145 cells using fluorescence-activated cell sorting (FACS). We used Illumina HumanHT-12 v4 Expression to investigate gene expression profiling of CSCs and non-CSCs. Protein-protein interaction (PPI) network analysis was performed using the STRING database. Biomarkers selected based on gene expression profiling were visually analyzed using immunofluorescence staining method. An image analysis program, ImageJ®, was used for the analysis of fluorescence intensity. In microarray analysis, we found that many ribosomal proteins and translation initiation factors that constitute the mTOR complex were highly expressed. PPI analysis using the 33 genes demonstrated that there was a close interaction between ribosome biogenesis, translation, and mTOR signaling. The fluorescence amount of mTOR and MLST8 were higher in CSCs compared to non-CSCs. The increase in a number of genes associated with ribosome biogenesis, translation, and mTOR signaling may be important to evaluate prognosis and determine treatment approach for prostate cancer (PCa). A better understanding of the molecular pathways associated with CSCs may be promising to develop targeted therapies to prolong survival in PCa.

Assembly of an atrazine catabolic operon and its introduction to Gram-negative hosts for robust and stable degradation of triazine herbicides.

In 1995, Pseudomonas sp. ADP, capable of metabolizing atrazine, was isolated from contaminated soil. Genes responsible for atrazine mineralization were found scattered in the 108.8 kb pADP-1 plasmid carried by this strain, some of them flanked by insertion sequences rendering them unstable. The goal of this work was to construct a transcriptional unit containing the atz operon in an easy to transfer manner, to be introduced and inherited stably by Gram-negative bacteria. atz genes were PCR amplified, joined into an operon and inserted onto the mobilizable plasmid pBAMD1-2. Primers were designed to add efficient transcription and translation signals. Plasmid bearing the atz operon was transferred to different Gram-negative strains by conjugation, which resulted in Tn5 transposase-mediated chromosomal insertion of the atz operon. To test the operon activity, atrazine degradation by transposants was assessed both colorimetrically and by high-performance liquid chromatography (HPLC). Transposants mineralized atrazine more efficiently than wild-type Pseudomonas sp. ADP and did not accumulate cyanuric acid. Atrazine degradation was not repressed by simple nitrogen sources. Genes conferring atrazine-mineralizing capacities were stable and had little or null effect on the fitness of different transposants. Introduction of catabolic operons in a stable fashion could be used to develop bacteria with better degrading capabilities useful in bioremediation.

EP.23A novel LMNA mutation identified in an Argentinian patient with autosomal dominant Emery-Dreifuss muscular dystrophy phenotype

LMNA mutations result in a broad array of diseases, named laminopathies. The Emery-Dreifuss muscular dystrophy and the dilated cardiomyopathy with conduction defects phenotypes, result primarily from heterozygous missense mutations in the LMNA gene (often in a dominant negative fashion), whilst truncating mutations resulting in premature termination codon are less common, but also described. We report a 38-year-old man with slowly progressive weakness and wasting in an humeroperoneal distribution beginning in early childhood, accompanied by elbow and achilles tendon contractures. At the age of 37, an implantable cardioverter defibrillator was required as he developed severe cardiac conduction defects. Incipient structural myocardial changes were also evident in the cardiac nuclear magnetic resonance. His family history reveals the existence of a twin sister with similar weakness pattern and contractures, without conduction or structural cardiac abnormalities. Additionally, both his father and paternal uncle suffered from dilated cardiomyopathy, without muscular weakness, and passed away from sudden death at 45 and 67 years of age respectively. Suspecting a Laminopathy a target next generation panel for cardiomyopathy and skeletal muscle disease was performed. A novel pathogenic variant c.210del (p.Ser71Alafs*25), at heterozygous state, was identified in LMNA. This sequence change creates a premature translational stop signal in the LMNA gene. It is expected to result in an absent or disrupted protein product. Loss-of-function variants in LMNA are known to be pathogenic. This variant is not present in population databases, and has not been reported in the literature in individuals with LMNA-related conditions. We present a 38-year-old patient with autosomal dominant Emery-Dreifuss muscular dystrophy phenotype, with strong family history, and a novel truncating variant identified in LMNA. Functional protein studies would contribute to confirm the pathogenicity of this variant.

Tuberculous meningitis in children is characterized by compartmentalized immune responses and neural excitotoxicity

Tuberculous meningitis (TBM) is the most severe form of TB with high rates of mortality and morbidity. Here we conduct RNA-sequencing on whole blood as well as on ventricular and lumbar cerebrospinal fluid (CSF) of pediatric patients treated for TBM. Differential transcript expression of TBM cases are compared with healthy controls in whole blood and with non-TB cerebral infection controls in CSF. Whole blood RNA-Seq analysis demonstrates a distinct immune response pattern in TBM, with significant increase in both canonical and non-canonical inflammasome activation and decrease in T-cell activation. In ventricular CSF, a significant enrichment associated with neuronal excitotoxicity and cerebral damage is detected in TBM. Finally, compartmental comparison in TBM indicates that the ventricular profile represents brain injury whereas the lumbar profile represents protein translation and cytokine signaling. Together, transcriptomic analysis shows that disease processes differ between the periphery and the central nervous system, and within brain compartments.

Spatial deictics in translation

AbstractRecent research on deictic elements has made extensive use of translation corpora to demonstrate that asymmetries between translational paradigms signal subtle contrasts from a systemic and pragmatic perspective. The purpose of the present study is to identify translation correspondences of the English spatial demonstrativethisin Lithuanian and to discuss the contrastive features within the analytical framework of translational explicitation and implicitation. The results indicate that in the prototypical situational usage there is a high degree of correspondence betweenthisand its Lithuanian translation, while textual uses demonstrate low formal equivalence. A tendency to implicitate stems from the null subject construction and object ellipsis in Lithuanian and is also a result of the availability of facultative deictic words in Lithuanian. The apparent systemic difference between the deictic systems of English and Lithuanian indicates a tendency towards optional implicitation and stylistic variation in literary translation.

Design of Low Power and Low Phase Noise Current Starved Ring Oscillator for RFID Tag EEPROM

Power dissipation of CMOS IC is a key factor in low power applications especially in RFID tag memories. Generally, tag memories like electrically erasable programmable read-only memory (EEPROM) require an internal clock generator to regulate the internal voltage level properly. In EEPROM, oscillator circuit can generate any periodic clock signal for frequency translation. Among different types of oscillators, a current starved ring oscillator (CSRO) is described in this research due to its very low current biasing source, which in turn restrict the current flows to reduce the overall power dissipation. The designed CSRO is limited to three stages to reduce the power dissipation to meet the specs. The simulated output shows that, the improved CSRO dissipates only 4.9 mW under the power supply voltage (VDD) 1.2 V in Silterra 130 nm CMOS process. Moreover, this designed oscillator has the lowest phase noise -119.38 dBc/Hz compared to other research works. In addition, the designed CSRO is able to reduce the overall chip area, which is only 0.00114 mm2. Therefore, this proposed low power and low phase noise CSRO will be able to regulate the voltage level successfully for low power RFID tag EEPROM.

Angular vestibuloocular reflex responses in Otop1 mice. I. Otolith sensor input is essential for gravity context-specific adaptation

The role of the otoliths in mammals in the angular vestibuloocular reflex (VOR) has been difficult to determine because there is no surgical technique that can reliably ablate them without damaging the semicircular canals. The Otopetrin1 (Otop1) mouse lacks functioning otoliths because of failure to develop otoconia but seems to have otherwise normal peripheral anatomy and neural circuitry. By using these animals we sought to determine the role of the otoliths in angular VOR baseline function and adaptation. In six Otop1 mice and six control littermates we measured baseline ocular countertilt about the three primary axes in head coordinates; baseline horizontal (rotation about an Earth-vertical axis parallel to the dorsal-ventral axis) and vertical (rotation about an Earth-vertical axis parallel to the interaural axis) sinusoidal (0.2-10 Hz, 20-100°/s) VOR gain (= eye/head velocity); and the horizontal and vertical VOR after gain-increase (1.5×) and gain-decrease (0.5×) adaptation training. Countertilt responses were significantly reduced in Otop1 mice. Baseline horizontal and vertical VOR gains were similar between mouse types, and so was horizontal VOR adaptation. For control mice, vertical VOR adaptation was evident when the testing context, left ear down (LED) or right ear down (RED), was the same as the training context (LED or RED). For Otop1 mice, VOR adaptation was evident regardless of context. Our results suggest that the otolith translational signal does not contribute to the baseline angular VOR, probably because the mouse VOR is highly compensatory, and does not alter the magnitude of adaptation. However, we show that the otoliths are important for gravity context-specific angular VOR adaptation. NEW & NOTEWORTHY This is the first study examining the role of the otoliths (defined here as the utricle and saccule) in adaptation of the angular vestibuloocular reflex (VOR) in an animal model in which the otoliths are reliably inactivated and the semicircular canals preserved. We show that they do not contribute to adaptation of the normal angular VOR. However, the otoliths provide the main cue for gravity context-specific VOR adaptation.

The influence of different types of translational inaccuracies on the genetic code structure

BackgroundThe standard genetic code is a recipe for assigning unambiguously 21 labels, i.e. amino acids and stop translation signal, to 64 codons. However, at early stages of the translational machinery development, the codons did not have to be read unambiguously and the early genetic codes could have contained some ambiguous assignments of codons to amino acids. Therefore, the goal of this work was to obtain the genetic code structures which could have evolved assuming different types of inaccuracy of the translational machinery starting from unambiguous assignments of codons to amino acids.ResultsWe developed a theoretical model assuming that the level of uncertainty of codon assignments can gradually decrease during the simulations. Since it is postulated that the standard code has evolved to be robust against point mutations and mistranslations, we developed three simulation scenarios assuming that such errors can influence one, two or three codon positions. The simulated codes were selected using the evolutionary algorithm methodology to decrease coding ambiguity and increase their robustness against mistranslation.ConclusionsThe results indicate that the typical codon block structure of the genetic code could have evolved to decrease the ambiguity of amino acid to codon assignments and to increase the fidelity of reading the genetic information. However, the robustness to errors was not the decisive factor that influenced the genetic code evolution because it is possible to find theoretical codes that minimize the reading errors better than the standard genetic code.

Feasibility of using a fully immersive virtual reality system for kinematic data collection

Commercially-available Virtual Reality (VR) systems have the potential to be effective tools for simultaneous visual manipulation and kinematic data collection. Previously, these systems have been integrated with research-grade motion capture systems to provide both functionalities; however, they are yet to be used as stand-alone systems for kinematic data collection. The present study aimed to validate the HTC VIVE VR system for kinematic data collection by evaluating the accuracy of its position and orientation signals. The VIVE controller and tracker were each compared to a Polhemus Liberty magnetic tracking system sensor for angular and translational measurement error and signal drift. A sensor from each system was mounted to opposite ends of a rigid segment which was driven through fifty rotations and fifty translations. Mean angular errors for both the VIVE tracker and controller were below 0.4°. Mean translational error for both sensors was below 3 mm. Drift in the Liberty signal components was consistently lower than drift in VIVE components. However, all mean rotational drift measures were below 0.1° and all mean translational measures were below 0.35 mm. These data indicate that the HTC VIVE system has the potential to be a valid and reliable means of kinematic data collection. However, further investigation is necessary to determine the VIVE’s suitability for capturing extremely minute or high-volume movements.

The Analysis Of Motor Imagery And SSVEPs For the BCI Application

Many studies have shown that the electrical and magnetic fields generated during brain activities can produce certain signals. Some of these signals can be captured using electroencephalography, a detection tool involving mobile brainwave sensors whose use has matured and become affordable. The brain-computer interface (BCI) provides an alternative form of communication between the human and a system (computer or actuator) without any physical contact between them. There are many ways to evoke brain signals for translation into computer tasks, but the most popular are motor imagery and steady-state visual evoked potentials (SSVEPs). In this research, an offline analysis of motor imagery and SSVEPs based on BCI experiments that use electroencephalography (EEG) is reported. The results show that SSVEPs are more accurate and convenient than motor imagery, with errors of 15 percent and 35 percent, respectively.

Vestibular System and Self-Motion.

Detection of the state of self-motion, such as the instantaneous heading direction, the traveled trajectory and traveled distance or time, is critical for efficient spatial navigation. Numerous psychophysical studies have indicated that the vestibular system, originating from the otolith and semicircular canals in our inner ears, provides robust signals for different aspects of self-motion perception. In addition, vestibular signals interact with other sensory signals such as visual optic flow to facilitate natural navigation. These behavioral results are consistent with recent findings in neurophysiological studies. In particular, vestibular activity in response to the translation or rotation of the head/body in darkness is revealed in a growing number of cortical regions, many of which are also sensitive to visual motion stimuli. The temporal dynamics of the vestibular activity in the central nervous system can vary widely, ranging from acceleration-dominant to velocity-dominant. Different temporal dynamic signals may be decoded by higher level areas for different functions. For example, the acceleration signals during the translation of body in the horizontal plane may be used by the brain to estimate the heading directions. Although translation and rotation signals arise from independent peripheral organs, that is, otolith and canals, respectively, they frequently converge onto single neurons in the central nervous system including both the brainstem and the cerebral cortex. The convergent neurons typically exhibit stronger responses during a combined curved motion trajectory which may serve as the neural correlate for complex path perception. During spatial navigation, traveled distance or time may be encoded by different population of neurons in multiple regions including hippocampal-entorhinal system, posterior parietal cortex, or frontal cortex.

'Stop' in protein synthesis is modulated with exquisite subtlety by an extended RNA translation signal.

Translational stop codons, UAA, UAG, and UGA, form an integral part of the universal genetic code. They are of significant interest today for their underlying fundamental role in terminating protein synthesis, but also for their potential utilisation for programmed alternative translation events. In diverse organisms, UAA has wide usage, but it is puzzling that the high fidelity UAG is selected against and yet UGA, vulnerable to suppression, is widely used, particularly in those archaeal and bacterial genomes with a high GC content. In canonical protein synthesis, stop codons are interpreted by protein release factors that structurally and functionally mimic decoding tRNAs and occupy the decoding site on the ribosome. The release factors make close contact with the decoding complex through multiple interactions. Correct interactions cause conformational changes resulting in new and enhanced contacts with the ribosome, particularly between specific bases in the mRNA and rRNA. The base following the stop codon (fourth or +4 base) may strongly influence decoding efficiency, facilitating alternative non-canonical events like frameshifting or selenocysteine incorporation. The fourth base is drawn into the decoding site with a compacted stop codon in the eukaryotic termination complex. Surprisingly, mRNA sequences upstream and downstream of this core tetranucleotide signal have a significant influence on the strength of the signal. Since nine bases downstream of the stop codon are within the mRNA channel, their interactions with rRNA, and r-proteins may affect efficiency. With this understanding, it is now possible to design stop signals of desired strength for specific applied purposes.

Functional and structural characterization of the chikungunya virus translational recoding signals

Climate change and human globalization have spurred the rapid spread of mosquito-borne diseases to naïve populations. One such emerging virus of public health concern is chikungunya virus (CHIKV), a member of the Togaviridae family, genus Alphavirus CHIKV pathogenesis is predominately characterized by acute febrile symptoms and severe arthralgia, which can persist in the host long after viral clearance. CHIKV has also been implicated in cases of acute encephalomyelitis, and its vertical transmission has been reported. Currently, no FDA-approved treatments exist for this virus. Recoding elements help expand the coding capacity in many viruses and therefore represent potential therapeutic targets in antiviral treatments. Here, we report the molecular and structural characterization of two CHIKV translational recoding signals: a termination codon read-through (TCR) element located between the nonstructural protein 3 and 4 genes and a programmed -1 ribosomal frameshift (-1 PRF) signal located toward the 3' end of the CHIKV 6K gene. Using Dual-Luciferase and immunoblot assays in HEK293T and U87MG mammalian cell lines, we validated and genetically characterized efficient TCR and -1 PRF. Analyses of RNA chemical modification data with selective 2'-hydroxyl acylation and primer extension (SHAPE) assays revealed that CHIKV -1 PRF is stimulated by a tightly structured, triple-stem hairpin element, consistent with previous observations in alphaviruses, and that the TCR signal is composed of a single large multibulged hairpin element. These findings illuminate the roles of RNA structure in translational recoding and provide critical information relevant for design of live-attenuated vaccines against CHIKV and related viruses.

NEW PATHOGENIC VARIANT ON IL-7R GENE ASSOCIATED WITH SCID

Introduction T-B+NK+ Severe Combined Immunodeficiency (SCID) due to mutations in the genes encoding the IL-7 receptor represent about 10% of American SCID cases. We report a patient found to have a c.675C>A (p.Tyr225*) homozygous mutation in exon 5. This variant is not present in population databases, and has not been reported in the literature in individuals with IL-7 receptor disease. Case Description Our medical center received a referral for abnormal T-cell receptor excision circle (TREC) results on a newborn screen. The patient's mother had an uncomplicated pregnancy, and there was no consanguinity or family history concerning for a primary immune deficiency. Lymphocyte subsets revealed an absence of T cells, along with present, but diminished B-cells and Natural-Killer cells. The patient had an absent thymic shadow on chest X-ray. He was also found to have poor mitogenic responses. FISH for DiGeorge syndrome was negative for deletions in 22q11.2 and 22q13, and testing for ADA deficiency was negative. Genetic testing resulted in a c.675C>A (p.Tyr225*) homozygous mutation. The patient has had a confirmed infection with Rhinovirus. He is currently on bacterial and fungal prophylaxis, and receiving immune globulin while awaiting bone marrow transplantation. Discussion A homozygous mutation in IL7R, Exon 5, c.675C>A (p.Tyr225*) creates a premature translational stop signal in the IL7R gene, resulting in an absent or disrupted protein product. Loss of function variants of IL7R are known to be pathogenic. This case reinforces the need for researchers to functionally evaluate all mutations found in patients.

Description and initial characterization of metatranscriptomic nidovirus-like genomes from the proposed new family Abyssoviridae, and from a sister group to the Coronavirinae, the proposed genus Alphaletovirus

Transcriptomics has the potential to discover new RNA virus genomes by sequencing total intracellular RNA pools. In this study, we have searched publicly available transcriptomes for sequences similar to viruses of the Nidovirales order. We report two potential nidovirus genomes, a highly divergent 35.9 kb likely complete genome from the California sea hare Aplysia californica, which we assign to a nidovirus named Aplysia abyssovirus 1 (AAbV), and a coronavirus-like 22.3 kb partial genome from the ornamented pygmy frog Microhyla fissipes, which we assign to a nidovirus named Microhyla alphaletovirus 1 (MLeV). AAbV was shown to encode a functional main proteinase, and a translational readthrough signal. Phylogenetic analysis suggested that AAbV represents a new family, proposed here as Abyssoviridae. MLeV represents a sister group to the other known coronaviruses. The importance of MLeV and AAbV for understanding nidovirus evolution, and the origin of terrestrial nidoviruses are discussed.

Cholesterol depletion enhances TGF-β Smad signaling by increasing c-Jun expression through a PKR-dependent mechanism.

Transforming growth factor-β (TGF-β) plays critical roles in numerous physiological and pathological responses. Cholesterol, a major plasma membrane component, can have pronounced effects on signaling responses. Cells continually monitor cholesterol content and activate multilayered transcriptional and translational signaling programs, following perturbations to cholesterol homeostasis (e.g., statins, the commonly used cholesterol-reducing drugs). However, the cross-talk of such programs with ligand-induced signaling responses (e.g., TGF-β signaling) remained unknown. Here, we studied the effects of a mild reduction in free (membrane-associated) cholesterol on distinct components of TGF-β–signaling pathways. Our findings reveal a new regulatory mechanism that enhances TGF-β–signaling responses by acting downstream from receptor activation. Reduced cholesterol results in PKR-dependent eIF2α phosphorylation, which enhances c-Jun translation, leading in turn to higher levels of JNK-mediated c-Jun phosphorylation. Activated c-Jun enhances transcription and expression of Smad2/3. This leads to enhanced sensitivity to TGF-β stimulation, due to increased Smad2/3 expression and phosphorylation. The phospho/total Smad2/3 ratio remains unchanged, indicating that the effect is not due to altered receptor activity. We propose that cholesterol depletion induces overactivation of PKR, JNK, and TGF-β signaling, which together may contribute to the side effects of statins in diverse disease settings.