Eukaryotic Elongation Factor 1A Research Articles

Cpmer: A new conserved eEF1A2-binding partner that regulates Eomes translation and cardiomyocyte differentiation.

SummaryPrevious studies have shown that eukaryotic elongation factor 1A2 (eEF1A2) serves as an essential heart-specific translation elongation element and that its mutation or knockout delays heart development and causes congenital heart disease and death among species. However, the function and regulatory mechanisms of eEF1A2 in mammalian heart development remain largely unknown. Here we identified the long noncoding RNA (lncRNA) Cpmer (cytoplasmic mesoderm regulator), which interacted with eEF1A2 to co-regulate differentiation of mouse and human embryonic stem cell-derived cardiomyocytes. Mechanistically, Cpmer specifically recognized Eomes mRNA by RNA-RNA pairing and facilitated binding of eEF1A2 with Eomes mRNA, guaranteeing Eomes mRNA translation and cardiomyocyte differentiation. Our data reveal a novel functionally conserved lncRNA that can specifically regulate Eomes translation and cardiomyocyte differentiation, which broadens our understanding of the mechanism of lncRNA involvement in the subtle translational regulation of eEF1A2 during mammalian heart development.

Computational Prediction of the Potential Target of SARS-CoV-2 Inhibitor Plitidepsin via Molecular Docking, Dynamic Simulations and MM-PBSA Calculations.

Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) replication depends on the interaction between the viral proteins and the human translation machinery. The cytotoxic peptide plitidepsin was found to inhibit CoV‐2 up to 90 % at a concentration of 0.88 nM. In vitro studies suggest that this activity may be attributed to the inhibition of the eukaryotic translation elongation factor 1A (eEF1A). However, recent reports raised the potential for other cellular targets which plitidepsin may use to exert its potent antiviral activity. The lack of data about these potential targets represents a major limitation for its structural optimization. This work describes the use of a molecular modeling approach to rationalize the in vitro antiviral activity of plitidepsin and to identify potential cellular targets. The developed protocol involves an initial molecular docking step followed by molecular dynamics and binding free energy calculations. The results reveal the potential for plitidepsin to bind to the active site of the key enzyme SARS‐CoV‐2 RdRp. The results also highlight the importance of van der Waals interactions for proper binding with the enzyme. We believe that the results presented in this study could provide the grounds for the optimization of plitidepsin analogs as SARS‐CoV‐2 inhibitors.

Identification of Novel Biomarkers in Platelets for Diagnosing Parkinson’s Disease

Background: Parkinson’s disease (PD) is a common neurodegenerative disease affecting the elderly, but there is no blood test for PD diagnosis in the clinic currently. This study aimed to explore promising biomarkers in platelets (PLTs) for PD diagnosis. Methods: PLTs were isolated from whole blood samples of PD patients and healthy controls (HCs), and RNA was extracted for sequencing. RNA-seq was performed on the Illumina HiSeq platform. Results: A total of 2,221 genes with differential transcript levels (GDTLs) were identified between PD patients and HCs, 1,041 of which are upregulated genes and 1,180 of which are downregulated genes. WASH5P was the most upregulated gene and AC114491.1 was the most downregulated gene. Among the top 12 most relevant genes, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), eukaryotic elongation factor 1A (EEF1A1), and cathepsin S (CTSS) were reported to be associated with PD. Furthermore, gene ontology analysis showed that the most significant term in biological processes was neutrophil degranulation; the most enriched term in cellular components was cytoplasmic vesicle lumen; and tumor necrosis factor receptor superfamily binding was the most significant term in molecular functions. In the Kyoto Encyclopedia of Genes and Genomes enrichment analysis, inflammation-related pathway accounts for the majority. Conclusion: Our findings demonstrated WASH5P, MALAT1, EEF1A1, and CTSS may be promising biomarkers in PD, which may contribute to improving the effectiveness and accuracy of diagnosis for PD in the future.

An Isoform of the Eukaryotic Translation Elongation Factor 1A (eEF1a) Acts as a Pro-Viral Factor Required for Tomato Spotted Wilt Virus Disease in Nicotiana benthamiana

The tripartite genome of the negative-stranded RNA virus Tomato spotted wilt orthotospovirus (TSWV) is assembled, together with two viral proteins, the nucleocapsid protein and the RNA-dependent RNA polymerase, into infectious ribonucleoprotein complexes (RNPs). These two viral proteins are, together, essential for viral replication and transcription, yet our knowledge on the host factors supporting these two processes remains limited. To fill this knowledge gap, the protein composition of viral RNPs collected from TSWV-infected Nicotiana benthamiana plants, and of those collected from a reconstituted TSWV replicon system in the yeast Saccharomyces cerevisiae, was analysed. RNPs obtained from infected plant material were enriched for plant proteins implicated in (i) sugar and phosphate transport and (ii) responses to cellular stress. In contrast, the yeast-derived viral RNPs primarily contained proteins implicated in RNA processing and ribosome biogenesis. The latter suggests that, in yeast, the translational machinery is recruited to these viral RNPs. To examine whether one of these cellular proteins is important for a TSWV infection, the corresponding N. benthamiana genes were targeted for virus-induced gene silencing, and these plants were subsequently challenged with TSWV. This approach revealed four host factors that are important for systemic spread of TSWV and disease symptom development.

Expression and Clinical Value of Eukaryotic Translation Elongation Factor 1A1 (EEF1A1) in Diffuse Large B Cell Lymphoma.

BackgroundThe eukaryotic translation elongation factor 1A1 (EEF1A1) participates in protein translation and has been reported to be involved in tumor progression such as hepatocellular carcinoma. Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoid malignancy in adults. In the present study, we aimed to detect the expression of EEF1A1 in DLBCL and to analyze its relationship with prognosis.MethodsWe reviewed medical records of DLBCL patients in our hospital and evaluated their expression level of EEF1A1 in tumor tissues using immunohistochemical (IHC) assay. The Chi-square method was used for correlation analysis. The Kaplan–Meier method with Log rank test was used for univariate analysis. Cox proportional hazards model was used for multivariate analysis. Cellular and mice models were introduced to validate its oncogenic role.ResultsEEF1A1 expression in tumor cells was higher in certain DLBCL cases. Patients with higher EEF1A1 expression were more likely to have advanced tumor stage and poorer 5-year overall survival (OS) rates. EEF1A1 expression in tumor cells was an independent risk predictor for OS (P < 0.05). Cellular assays demonstrated that EEF1A1-shRNA significantly inhibited lymphoma cell proliferation. The study of xenografts further verified the effect of EEF1A1-shRNA on suppressing tumor growth in vivo.ConclusionEEF1A1 positivity predicts short survival in DLBCL patients. For patients with higher EEF1A1 expression, more strategy such as anti-EEF1A1 antibody treatment should be developed.

MLIF Modulates Microglia Polarization in Ischemic Stroke by Targeting eEF1A1.

Monocyte locomotion inhibitory factor (MLIF) is a heat-stable pentapeptide from Entamoeba histolytica. Our previous study found that MLIF protects against ischemic stroke in rats and mice and exerts a neuroprotection effect in human neuroblastoma SH-SY5Y cells. Microglia/macrophage polarization has been proven to be vital in the pathology of ischemic stroke. Nevertheless, whether MLIF is able to modulate microglia/macrophage polarization remains unclear. We performed middle cerebral artery occlusion (MCAO) on C57BL/6J male mice and induced cultured BV2 microglia by oxygen-glucose deprivation (OGD), respectively. Immunfluorescence was utilized to detect the M1/2 markers, such as CD206 and CD16/32. qPCR and ELISA were used to detect the signature gene change of M1/2. The MAPK and NF-κB pathway associated proteins were measured by Western blot. To identify the protein target of MLIF, a pull-down assay was performed. We found that MLIF promoted microglia transferring from a “sick” M1 phenotype to a “healthy” M2 phenotype in vivo or in vitro. Furthermore, we proved that eukaryotic elongation factor 1A1 (eEF1A1) was involved in the modulation of microglia/macrophage polarization. Knocking down eEF1A1 by siRNA exhibited the M1 promotion effect and M2 inhibition effect. Taken together, our results demonstrated MLIF modulated microglia/macrophage polarization by targeting eEF1A1 in ischemic stroke.

Nannocystin ax, an eEF1A inhibitor, induces G1 cell cycle arrest and caspase-independent apoptosis through cyclin D1 downregulation in colon cancer in vivo

Colorectal cancer (CRC) is one of the most common causes of cancer-related death worldwide. Nannocystin ax (NAN), a 21-membered cyclodepsipeptide initially isolated from myxobacteria of the Nannocystis genus, was found to target the eukaryotic elongation factor 1A (eEF1A). The current study was designed to evaluate the anticancer effect and underlying mechanisms of NAN with in vitro and in vivo models. Results showed that NAN induced G1 phase cell cycle arrest and caspase-independent apoptosis in HCT116 and HT29 human CRC cells. NAN significantly downregulated cyclin D1 level in a short time, but NAN did not affect the transcription level and ubiquitin-dependent degradation of cyclin D1. Furthermore, NAN treatment directly targeted eEF1A and partially decreased the synthesis of new proteins, contributing to the downregulation of cyclin D1. Besides, NAN significantly suppressed tumor growth in the zebrafish xenograft model. In conclusion, NAN triggered G1 phase cell cycle arrest through cyclin D1 downregulation and eEF1A-targeted translation inhibition and promoted caspase-independent apoptosis in CRC cells.

METTL21B is a prognostic biomarker and potential therapeutic target in low-grade gliomas.

A considerable amount of literature has demonstrated that eukaryotic translation elongation factor 1A (eEF1A) is closely related to tumors. As a newly identified lysine specific methyltransferase targeting eEF1A at Lys-165, too little attention has been paid to the function of METTL21B. To determine the potential significance and prognostic value of METTL21B in low grade glioma (LGG), we analyzed the expression, methylation level and copy number variations (CNV) of METTL21B and its effect on prognosis in patients with LGG by 4 public databases in conjunction with experimental examination of LGG patient samples. As a result, we found that high expression, hypomethylation and gain/amplification of CNV of METTL21B were associated with poor prognosis in LGG. The potential functions of METTL21B in LGG may be involved in cell adhesion, angiogenesis and cell proliferation of tumor by enrichment analysis. In addition, METTL21B may facilitate immune evasion of tumor and affect prognosis by mediating macrophage polarization from M1 to M2 and regulating expression of immune checkpoints. Nevertheless, patients with high METTL21B level are likely to have better response to immune checkpoints blockage therapy. Because of its substrate specificity, METTL21B is expected to be a promising target for the treatment of glioma.

Mutational analysis reveals potential phosphorylation sites in eukaryotic elongation factor 1A that are important for its activity.

Previous studies have suggested that phosphorylation of translation elongation factor 1A (eEF1A) can alter its function, and large‐scale phospho‐proteomic analyses in Saccharomyces cerevisiae have identified 14 eEF1A residues phosphorylated under various conditions. Here, a series of eEF1A mutations at these proposed sites were created and the effects on eEF1A activity were analyzed. The eEF1A‐S53D and eEF1A‐T430D phosphomimetic mutant strains were inviable, while corresponding alanine mutants survived but displayed defects in growth and protein synthesis. The activity of an eEF1A‐S289D mutant was significantly reduced in the absence of the guanine nucleotide exchange factor eEF1Bα and could be restored by an exchange‐deficient form of the protein, suggesting that eEF1Bα promotes eEF1A activity by a mechanism other than nucleotide exchange. Our data show that several of the phosphorylation sites identified by high‐throughput analysis are critical for eEF1A function.

Identifying the Cellular Target of Cordyheptapeptide A and Synthetic Derivatives.

Cordyheptapeptide A is a lipophilic cyclic peptide from the prized Cordyceps fungal genus that shows potent cytotoxicity in multiple cancer cell lines. To better understand the bioactivity and physicochemical properties of cordyheptapeptide A with the ultimate goal of identifying its cellular target, we developed a solid-phase synthesis of this multiply N-methylated cyclic heptapeptide which enabled rapid access to both side chain- and backbone-modified derivatives. Removal of one of the backbone amide N-methyl (N-Me) groups maintained bioactivity, while membrane permeability was also preserved due to the formation of a new intramolecular hydrogen bond in a low dielectric solvent. Based on its cytotoxicity profile in the NCI-60 cell line panel, as well as its phenotype in a microscopy-based cytological assay, we hypothesized that cordyheptapeptide was acting on cells as a protein synthesis inhibitor. Further studies revealed the molecular target of cordyheptapeptide A to be the eukaryotic translation elongation factor 1A (eEF1A), a target shared by other lipophilic cyclic peptide natural products. This work offers a strategy to study and improve cyclic peptide natural products while highlighting the ability of these lipophilic compounds to effectively inhibit intracellular disease targets.

On the Need to Tell Apart Fraternal Twins eEF1A1 and eEF1A2, and Their Respective Outfits.

eEF1A1 and eEF1A2 are paralogous proteins whose presence in most normal eukaryotic cells is mutually exclusive and developmentally regulated. Often described in the scientific literature under the collective name eEF1A, which stands for eukaryotic elongation factor 1A, their best known activity (in a monomeric, GTP-bound conformation) is to bind aminoacyl-tRNAs and deliver them to the A-site of the 80S ribosome. However, both eEF1A1 and eEF1A2 are endowed with multitasking abilities (sometimes performed by homo- and heterodimers) and can be located in different subcellular compartments, from the plasma membrane to the nucleus. Given the high sequence identity of these two sister proteins and the large number of post-translational modifications they can undergo, we are often confronted with the dilemma of discerning which is the particular proteoform that is actually responsible for the ascribed biochemical or cellular effects. We argue in this review that acquiring this knowledge is essential to help clarify, in molecular and structural terms, the mechanistic involvement of these two ancestral and abundant G proteins in a variety of fundamental cellular processes other than translation elongation. Of particular importance for this special issue is the fact that several de novo heterozygous missense mutations in the human EEF1A2 gene are associated with a subset of rare but severe neurological syndromes and cardiomyopathies.

Correlation of elongation factor 1A accumulation with photosynthetic pigment content and yield in winter wheat varieties under heat stress conditions

Heat stress is one of the most important environmental factors that influences wheat growth and development, leading to significant losses in grain yield and has become a significant detrimental factor for worldwide wheat production. In recent years, several studies suggested that eukaryotic elongation factor 1A (eEF1A), may contribute to heat tolerance in plants, therefore the aim of this study was: to investigate the accumulation of eEF1A in wheat under conditions of moderate and high air temperatures; to determine the amount of photosynthetic pigments and to determine the yield traits; and to examine whether there is a correlation between eEF1A accumulation, photosynthetic pigments, and yield in different wheat varieties. The results showed that heat stress induced accumulation of eEF1A significantly different among wheat varieties and showed that varieties with a higher accumulation of eEF1A under heat stress are characterized by a smaller decrease in the photosynthetic pigments. A correlation between higher accumulation of eEF1A under heat stress and yield traits was found. Analyzed parameters from two growing seasons, indicated that the higher accumulation of eEF1A and a smaller decrease in photosynthetic pigments distinguishes the varieties more resistant to heat stress. The analysis of the molecular mechanisms by immunoblot, under conditions of high and moderate air temperatures in two growing seasons, aims to develop agricultural strategy and develop wheat varieties tolerant to heat stress.

Inhibition of Elongation Factor 1A1 Activity Decreases Lipid Droplet Accumulation

Non-alcoholic fatty liver disease (NAFLD) is characterized by hepatocyte lipid accumulation, which can cause hepatocyte injury through lipotoxicity. Eukaryotic elongation factor 1A1 (EEF1A1), which is a component of the protein synthetic machinery and regulates actin cytoskeleton turnover, is induced in human hepatocyte-like HepG2 cells undergoing lipotoxicity, in association with ER stress. Moreover, EEF1A1 has been identified in the lipid droplet (LD) proteome in various cell and tissue types, suggesting that it may have a role in LD biology. Partial inhibition of EEF1A1 peptide elongation activity with the marine compound didemnin B (DB) decreases lipotoxic HepG2 cell death. However, the effects of EEF1A1 inhibition on cellular lipid accumulation are unclear. Based on the potential association of EEF1A1 with LDs, and its apparent roles in responding to ER stress and in regulating cytoskeleton dynamics, we hypothesized that inhibition of EEF1A1 with DB would alter hepatocyte LD accumulation. Male 129S6 mice were fed a western diet for 26 weeks to induce NAFLD, followed by intervention with DB (50 μg/kg, i.p.) once every 3 days for 14 days. Treatment with DB decreased liver lipid accumulation, as determined by biochemical and histological analyses, and upregulated expression of genes involved in fatty acid oxidation (Cpt1a, Fgf21). Assessments of LD size in mouse liver revealed a significant decrease in median size upon treatment with DB, suggesting that DB promotes the formation of smaller LDs. To explore whether this effect was conserved in human hepatocyte-like cells, HepG2 cells were treated for 6 h with DB (80 nM) under lipotoxic conditions (1 mM palmitate plus oleate) and LDs were visualized by confocal microscopy. Cells exposed to lipotoxic conditions exhibited increased LD area, and treatment with DB appeared to decrease the formation of large LDs (-63%, p=0.11). This occurred alongside upregulation of CPT1A mRNA, which was consistent with the upregulation of genes involved in fatty acid oxidation observed in mouse liver. Finally, a Chinese hamster ovary cell line that harbors a mutation leading to reduced EEF1A1 expression accumulated less triglyceride under high fatty acid conditions compared to wild-type cells. Together, these findings suggest that that disruption of EEF1A1 activity may diminish LD accumulation and channel excess fatty acids toward oxidation, which could have implications for the treatment of NAFLD.

Selection of Reference Genes for RT-qPCR Studies in Different Organs of Rice Cultivar BRS AG Submitted to Recurrent Saline Stress

Quantitative real-time polymerase chain reactions (RT-qPCR) have become one of the most widely used methods for analyzing gene expression, provided suitable reference genes are available to normalize the data. RNA was isolated from leaves, grain, rachises and sheaths of rice (Oryza sativa L. cv. BRS AG) submitted to different saline stress events for seven days, and expression analysis was carried out by RT-qPCR. Expression levels of ten candidate reference genes were assessed, actin11 (ACT11), ubiquitin conjugating enzyme E2 (UBC-E2), eukaryotic elongation factor1-α (Eef-1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin (β-Tub), eukaryotic initiation factor 4a (Eif-4-α), ubiquitin10 (UBQ10), ubiquitin5 (UBQ5), aquaporin TIP41 (TIP41-like). Gene expression stability was calculated using the common statistical algorithms geNorm, BestKeeper and ΔCt method, NormFinder and RefFinder. The most stably expressed genes were UBC2E and GAPDH for leaves, UBQ5 and UBQ10 for sheaths, TIP41 and UBQ10 for rachises, and TIP41 and cyclophilin for grain. Gene expression of triose phosphate translocator (TPT1), ADP-glucose transporter (BT1-1), choline monooxygenase (CMO) was used to validate the selected reference genes. The results highlighted the importance of using suitable reference gene to normalize gene expression data in rice plants.

A pivotal study in human prostate cancer tissues and cell lines to measure the expression levels of eukaryotic Elongation Factor 1A proteins and the effect of a nucleic acid-based GT75 aptamer

Background: Two major isoforms of the eukaryotic Elongation Factor 1A are recognised: the eEF1A1, ubiquitously expressed, and the eEF1A2 presents in adult heart and skeletal muscle, in nervous system and in some other specialized cells. Both proteins are implicated in the development and progression of human cancers with different role. In prostate cancer, overexpression of eEF1A2 has been proposed to be involved in the onset of the tumour and it was related to a worse outcome. However, the expression level of both proteins in prostate cancer tissue is not well known. We explore the expression level of the two proteins and mRNAs in formalin-fixed paraffin-embedded tissues derived from prostate cancer patients. Moreover, we investigated the effect of the nucleic acid-based GT75 aptamer, targeting eEF1A protein, in human prostate cancer cell lines.

S1352 Dimethylation Level of eEF1A and the Gastric Mucosa After Helicobacter pylori (H. pylori) Eradication Therapy

INTRODUCTION: The eradication therapy of H. pylori infections removes the primary cause of gastric cancer (GC). However, the effectiveness for prevention of GC depends on the severity and extent of atrophic damage. Therefore, even after of eradication therapy, it is important to identify patients at high-risk for gastric cancer. Methylation accumulation which is caused by exposure to chronic inflammation, such as H. pylori infection, is known to be associated with increased risk of GC. Recent study showed that the dimethylation of eukaryotic elongation factor 1A (eEF1A) lead to upregulation of activity and promote carcinogenesis in vivo. In this study, we investigate the relationship between the dimethylation of eEF1A, that active GTPase form, associated with the clinical factor, histological status based on cell type, and GC in the H. pylori absent gastric mucosa after eradication therapy. METHODS: Records of 72 post-H. pylori eradication patients who underwent upper gastrointestinal endoscopy between 2001 and 2018 retrospectively reviewed. HE staining performed to assess the degrees of inflammation and atrophy a were scored according to the Updated Sydney System. We evaluate the eEF1A demethylation levels using anti-eEF1AK55me2 (demethylation of eEF1A) in each functional types of cells (surface mucous cells, parietal cells, mucous neck cells and chief cells) in fundic gland. Additionally, we investigated the relationship between the gastric cancer risk, clinical data and eEF1A dimethylation levels using univariate and multivariate analyses. RESULTS: The dimethylation level of eEF1A associated with degrees of histological atrophy (P = 0.0404) and GC treatment history (P = 0.0002) in PG1 positive chief cell area, located at bottoms of fundic gland of post eradication patients. Univariate analyses showed that histological inflammation, atrophy (not endoscopic) and eEF1A dimethylation levels in bottoms of fundic gland. Multivariate analysis revealed that histological atrophic levels and highly-dimethylation of eEF1A in bottoms of mucosa (PG1 positive chief cell area) was the sole independent factor related to the diagnosis of GC (OR, 8.516; 95% CI, 1.217–59.6, P = 0.031). CONCLUSION: The demethylation level of eEF1A in bottom of mucosa associated with an increased risk of gastric cancer in after H. pylori eradication therapy. Further long-term prospective study will be required.

The marine compound and elongation factor 1A1 inhibitor, didemnin B, provides benefit in western diet-induced non-alcoholic fatty liver disease

Inhibition of eukaryotic elongation factor 1A1 (EEF1A1) with the marine compound didemnin B decreases lipotoxic HepG2 cell death in vitro and improves early stage non-alcoholic fatty liver disease (NAFLD) in young genetically obese mice. However, the effects of didemnin B on NAFLD in a model of long-term diet-induced obesity are not known. We investigated the effects of didemnin B on NAFLD severity and metabolic parameters in western diet-induced obese mice, and on the cell types that contribute to liver inflammation and fibrosis in vitro. Male 129S6 mice were fed either standard chow or western diet for 26 weeks, followed by intervention with didemnin B (50 μg/kg) or vehicle by intraperitoneal (i.p.) injection once every 3 days for 14 days. Didemnin B decreased liver and plasma triglycerides, improved oral glucose tolerance, and decreased NAFLD severity. Moreover, didemnin B moderately increased hepatic expression of genes involved in ER stress response (Perk, Chop), and fatty acid oxidation (Fgf21, Cpt1a). In vitro, didemnin B decreased THP-1 monocyte proliferation, disrupted THP-1 monocyte-macrophage differentiation, decreased THP-1 macrophage IL-1β secretion, and decreased hepatic stellate cell (HSteC) proliferation and collagen secretion under both basal and lipotoxic (high fatty acid) conditions. Thus, didemnin B improves hepatic steatosis, glucose tolerance, and blood lipids in obesity, in association with moderate, possibly hormetic, upregulation of pathways involved in cell stress response and energy balance in the liver. Furthermore, it decreases the activity of the cell types implicated in liver inflammation and fibrosis in vitro. These findings highlight the therapeutic potential of partial protein synthesis inhibition in the treatment of NAFLD.

Identification of proteins involved in transcription/translation (eEF 1A1) as an inhibitor of Bax induced apoptosis

Eukaryotic elongation factor 1A1 (eEF1A1) is central to translational activity. It is involved in complexes that form signal transduction with protein kinase C, as well as being a signal transducer and activator of transcription 3. eEF1A1 and eEF1A2 are isoforms of the alpha subunit of elongating factor 1 complex. It has been reported that eEF1A1 is expressed in most human tissues but the brain, skeletal muscle and heart. eEF1A1 has been linked to both apoptosis and anti-apoptotic activities. In this study, eEF1A1 was co-expressed with Bax, a proapoptotic protein via heterologous expression of recombinant DNA in yeast cells. Assays were carried out to monitor the fate and state of yeast cells when eEF1A1 was co-expressed with Bax. The yeast strain (bearing an integrated copy of the Bax gene) was transformed with an episomal 2-micron plasmid that encodes HA-tagged eEF1A1 gene. The resultant strain would allow co-expression of Bax and eEF1A1 in yeast cells, Bax being under the control of the GAL1 promoter, while the PGK1 promoter drives eEF1A1 expression. Bcl 2A1, a known anti-apoptotic protein, was also co-expressed with Bax in yeast cells as a positive control, to study the anti-apoptotic characteristic of eEF-1A1. The part eEF1A1 plays in apoptosis has been contentious, amidst the pro and anti-apoptotic properties of eEF1A1, it was shown clearly, in this study that eEF1A1 portrays only anti-apoptotic property in the presence of pro-apoptotic protein, Bax.

EEF1A1 deacetylation enables transcriptional activation of remyelination

Remyelination of the peripheral and central nervous systems (PNS and CNS, respectively) is a prerequisite for functional recovery after lesion. However, this process is not always optimal and becomes inefficient in the course of multiple sclerosis. Here we show that, when acetylated, eukaryotic elongation factor 1A1 (eEF1A1) negatively regulates PNS and CNS remyelination. Acetylated eEF1A1 (Ac-eEF1A1) translocates into the nucleus of myelinating cells where it binds to Sox10, a key transcription factor for PNS and CNS myelination and remyelination, to drag Sox10 out of the nucleus. We show that the lysine acetyltransferase Tip60 acetylates eEF1A1, whereas the histone deacetylase HDAC2 deacetylates eEF1A1. Promoting eEF1A1 deacetylation maintains the activation of Sox10 target genes and increases PNS and CNS remyelination efficiency. Taken together, these data identify a major mechanism of Sox10 regulation, which appears promising for future translational studies on PNS and CNS remyelination.

Control of translation elongation in health and disease.

ABSTRACTRegulation of protein synthesis makes a major contribution to post-transcriptional control pathways. During disease, or under stress, cells initiate processes to reprogramme protein synthesis and thus orchestrate the appropriate cellular response. Recent data show that the elongation stage of protein synthesis is a key regulatory node for translational control in health and disease. There is a complex set of factors that individually affect the overall rate of elongation and, for the most part, these influence either transfer RNA (tRNA)- and eukaryotic elongation factor 1A (eEF1A)-dependent codon decoding, and/or elongation factor 2 (eEF2)-dependent ribosome translocation along the mRNA. Decoding speeds depend on the relative abundance of each tRNA, the cognate:near-cognate tRNA ratios and the degree of tRNA modification, whereas eEF2-dependent ribosome translocation is negatively regulated by phosphorylation on threonine-56 by eEF2 kinase. Additional factors that contribute to the control of the elongation rate include epigenetic modification of the mRNA, coding sequence variation and the expression of eIF5A, which stimulates peptide bond formation between proline residues. Importantly, dysregulation of elongation control is central to disease mechanisms in both tumorigenesis and neurodegeneration, making the individual key steps in this process attractive therapeutic targets. Here, we discuss the relative contribution of individual components of the translational apparatus (e.g. tRNAs, elongation factors and their modifiers) to the overall control of translation elongation and how their dysregulation contributes towards disease processes.