Amino Acid Response Research Articles

Plasma Amino Acid Response in Mature Geldings Fed Three Concentrations of Lysine

Plasma Amino Acid Response in Mature Geldings Fed Three Concentrations of Lysine

BCL-2 Is a Downstream Target of ATF5 That Mediates the Prosurvival Function of ATF5 in a Cell Type-dependent Manner

ATF5 loss of function has been shown previously to cause apoptotic cell death in glioblastoma and breast cancer cells but not in non-transformed astrocytes and human breast epithelial cells. The mechanism for the cell type-dependent survival function of ATF5 is unknown. We report here that the anti-apoptotic factor BCL-2 is a downstream target of ATF5 that mediates the prosurvival function of ATF5 in C6 glioma cells and MCF-7 breast cancer cells. ATF5 binds to an ATF5-specific regulatory element that is downstream of and adjacent to the negative regulatory element in the BCL-2 P2 promoter, stimulating BCL-2 expression. Highlighting the critical role of BCL-2 in ATF5-dependent cancer cell survival, expression of BCL-2 blocks death of C6 and MCF-7 cells induced by dominant-negative ATF5, and depletion of BCL-2 impairs ATF5-promoted cell survival. Moreover, we found that BCL-2 expression is not regulated by ATF5 in non-transformed rat astrocytes, mouse embryonic fibroblasts, and human breast epithelial cells, where expression of BCL-2 but not ATF5 is required for cell survival. These findings identify BCL-2 as an essential mediator for the cancer-specific cell survival function of ATF5 in glioblastoma and breast cancer cells and provide direct evidence that the cell type-specific function of ATF5 derives from differential regulation of downstream targets by ATF5 in different types of cells.

Amino Acid Availability Controls TRB3 Transcription in Liver through the GCN2/eIF2α/ATF4 Pathway

In mammals, plasma amino acid concentrations are markedly affected by dietary or pathological conditions. It has been well established that amino acids are involved in the control of gene expression. Up to now, all the information concerning the molecular mechanisms involved in the regulation of gene transcription by amino acid availability has been obtained in cultured cell lines. The present study aims to investigate the mechanisms involved in transcriptional activation of the TRB3 gene following amino acid limitation in mice liver. The results show that TRB3 is up-regulated in the liver of mice fed a leucine-deficient diet and that this induction is quickly reversible. Using transient transfection and chromatin immunoprecipitation approaches in hepatoma cells, we report the characterization of a functional Amino Acid Response Element (AARE) in the TRB3 promoter and the binding of ATF4, ATF2 and C/EBPβ to this AARE sequence. We also provide evidence that only the binding of ATF4 to the AARE plays a crucial role in the amino acid-regulated transcription of TRB3. In mouse liver, we demonstrate that the GCN2/eIF2α/ATF4 pathway is essential for the induction of the TRB3 gene transcription in response to a leucine-deficient diet. Therefore, this work establishes for the first time that the molecular mechanisms involved in the regulation of gene transcription by amino acid availability are functional in mouse liver.

A Low-Protein Diet during Gestation in Rats Activates the Placental Mammalian Amino Acid Response Pathway and Programs the Growth Capacity of Offspring1–3

Placental efficiency is a predictor of fetal growth and development, which is also controlled by maternal gestational health and diet. The present study investigated the effects of a gestational low-protein diet on offspring growth capacity as well as the diet’s contribution to altered expression of placental genes associated with the mammalian amino acid response (AAR) pathway. To assess these outcomes, timed-pregnant Sprague Dawley rats were fed a control (C) diet with 18% protein or a low-protein (LP) diet with 9% protein throughout gestation (Expt. 1) or throughout gestation and lactation (Expt. 2). Placentas were collected during natural delivery and quantitative RT-PCR and Western-blot analyses were performed to determine placental mRNA and protein levels. By the end of the lactation period, offspring of dams fed the LP diet had stunted growth in both experiments. mRNA expression of target genes in the AAR pathway, such as activating transcription factor-3 (Atf3), asparagine synthetase (Asns), and Sodium-dependent neutral amino acid transporter-2 (Snat2), was greater in placentas of rats fed the LP diet compared with controls, as were placental ATF4 and p-eIF2α protein levels. The increase in mRNA expression of AAR pathway-associated genes was correlated with the stunting of offspring growth (Atf3: R2 = 0.32, P = 0.086; Asns: R2 = 0.44, P < 0.05; Snat2:R2 = 0.33, P = 0.084). Our study showed that the mammalian AAR pathway in placenta is upregulated by a maternal low-protein diet and this activation may act as a cue for the fetus to develop an adaptive response suited to their predicted postnatal environment, i.e. a more favorable phenotype for their survival.

Effects of leucine or whey protein addition to an oral glucose solution on serum insulin, plasma glucose and plasma amino acid responses in horses at rest and following exercise

Providing protein or amino acid mixtures in combination with glucose to post exercise in man has resulted in increases in the post feeding insulin response and in muscle glycogen and protein synthesis rates. However, whether protein and/or amino acids can modify the post exercise insulin responses in horses remains to be fully elucidated. To determine whether whey protein or leucine addition to a glucose solution affects the post gavage plasma insulin, glucose and amino acid responses in horses and whether these responses are different following a period of exercise vs. rest. Six mature, conditioned Thoroughbreds received a nasogastric gavage containing either 1 g/kg bwt glucose (G), G + 0.3 g/kg bwt whey protein (GW) or G + 0.3 g/kg bwt leucine (GL), following a period of either rest (R) or an exercise test on a high speed treadmill (EX). Each horse was studied under all 6 treatment conditions, separated by 10 day intervals. Blood samples were collected pre-exercise/rest, pregavage and at regular intervals up to 300 min post gavage. Plasma was analysed for glucose and amino acid concentrations and serum insulin concentrations were determined. There was a significantly (P < 0.05) greater insulin response in GL-R and GL-EX when compared to the other treatments. When compared to rest, post exercise plasma glucose responses were lower in G and GW but unchanged following GL administration. Plasma alanine concentrations were elevated post exercise in all EX treatments. With the exception of markedly elevated plasma leucine concentrations after GL-R and GL-EX, the plasma concentrations of all indispensable amino acids decreased during the post gavage period. Leucine but not whey protein augmented the serum insulin response to an oral glucose load. Leucine supplementation warrants further investigation as a means to increase the rate of post exercise muscle glycogen synthesis in horses.

Regulation of the human CHOP gene promoter by the stress response transcription factor ATF5 via the AARE1 site in human hepatoma HepG2 cells

Aims Activating transcription factor (ATF) 5 is a member of the cAMP response element-binding protein (CREB)/ATF family of transcription factors. We have shown that ATF5 is a stress response transcription factor that responds to amino acid limitation, arsenite exposure, or cadmium exposure. In this study we investigated whether ATF5 is involved in the regulation of CCAAT/enhancer-binding protein (C/EBP) homologous protein ( CHOP) gene expression. Main methods We used a transient transfection system to express ATF5 and analyzed the regulation of CHOP gene promoter in human hepatoma, HepG2 cells. We also studied the effect of ATF5 knockdown on arsenite-induced CHOP protein expression and arsenite-induced cell death of HepG2 cells. Key findings We showed that ATF5 activates the CHOP gene promoter in HepG2 cells. Both deletion analysis and point mutations of the promoter revealed that amino acid response element (AARE) 1 is responsible for ATF5-dependent promoter activation. Furthermore, the existence of either AARE1 or activating protein-1 (AP-1) site is sufficient for transcriptional activation of the CHOP gene promoter by arsenite exposure, although complete induction requires the existence of both elements. We also demonstrated that knockdown of ATF5 reduced arsenite-induced CHOP protein expression and arsenite-induced cell death of HepG2 cells. Significance These results suggested that the CHOP gene is a potential target for ATF5, and that ATF5 raises the arsenite-induced CHOP gene expression level via the AARE1 site in HepG2 cells.

Gene expression and integrated stress response in HepG2/C3A cells cultured in amino acid deficient medium.

The integrated stress response (ISR), a defense mechanism cells employ when under stress (e.g., amino acid deprivation), causes suppression of global protein synthesis along with the paradoxical increased expression of a host of proteins that are useful in combating various stresses. Genes that were similarly differentially expressed under conditions of either leucine- or cysteine-depletion were identified. Many of the genes known to contain an amino acid response element and to be induced in response to eIF2α phosphorylation and ATF4 heterodimer binding (ATF3, C/EBPβ, SLC7A1, SLC7A11, and TRIB3), as well as others shown to be induced downstream of eIF2α phosphorylation (C/EBPγ, CARS, SARS, CLCN3, CBX4, and PPP1R15A) were among the upregulated genes. Evidence for the induction of the ISR in these cells also included the increased phosphorylation of eIF2α and increased protein abundance of ATF4, ATF3, and ASNS in cysteine- and leucine-depleted cells. Based on genes highly differentially expressed in both leucine- and cysteine-deficient cells, a list of 67 downregulated and 53 upregulated genes is suggested as likely targets of essential amino acid deprivation in mammalian cells.

Maternal Low Protein Diet Induces Amino Acid Response (AAR) Pathway in the Liver of Dam Rat

ObjectiveAmino acid response (AAR) pathway reacts to deficiency of dietary protein or an imbalance of essential amino acids. The effects of low protein diet to dam rats during gestation have not been reported. The present study is designed to investigate the impacts of maternal diets on activation of the AAR pathway in dam rats.MethodsTimed‐pregnant Sprague‐Dawley rats were fed on one of the two isocaloric diets, control (20 kcal% protein, 64 kcal% carbohydrate) or low protein diet (8 kcal% protein, 76 kcal% carbohydrate) during gestation and lactation. The livers of dam rats were collected and used to isolate mRNA and protein.ResultsLow protein diet induces phosphorylation of eIF2alpha and ERK in the liver of dam rats. There were significantly changes in mRNA expression of the downstream target genes in the AAR pathway, such as ASNS (asparagine synthetase), ATF3 (activation transcription factor 3), CHOP(C/EBP homology protein), and SNAT2 (sodium‐coupled neutral amino acid transporter 2).ConclusionMaternal high carbohydrate/low‐protein diets fed during gestation and lactation regulates the activation of the AAR pathway in the livers of dam rats.Grant Funding Source: USDA

Expression profiling after activation of amino acid deprivation response in HepG2 human hepatoma cells

Dietary protein malnutrition is manifested as amino acid deprivation of individual cells, which activates an amino acid response (AAR) that alters cellular functions, in part, by regulating transcriptional and posttranscriptional mechanisms. The AAR was activated in HepG2 human hepatoma cells, and the changes in mRNA content were analyzed by microarray expression profiling. The results documented that 1,507 genes were differentially regulated by P < 0.001 and by more than twofold in response to the AAR, 250 downregulated and 1,257 upregulated. The spectrum of altered genes reveals that amino acid deprivation has far-reaching implications for gene expression and cellular function. Among those cellular functions with the largest numbers of altered genes were cell growth and proliferation, cell cycle, gene expression, cell death, and development. Potential biological relationships between the differentially expressed genes were analyzed by computer software that generates gene networks. Proteins that were central to the most significant of these networks included c-myc, polycomb group proteins, transforming growth factor β1, nuclear factor (erythroid-derived 2)-like 2-related factor 2, FOS/JUN family members, and many members of the basic leucine zipper superfamily of transcription factors. Although most of these networks contained some genes that were known to be amino acid responsive, many new relationships were identified that underscored the broad impact that amino acid stress has on cellular function.

Comparative proteomics analysis of differentially accumulated proteins in juice sacs of ponkan ( Citrus reticulata) fruit during postharvest cold storage

Comparative proteomics analysis was carried out in ‘Egan No.1’ ponkan ( Citrus reticulate cv. Egan No.1) fruit during low temperature storage. Commercially mature fruit were harvested, stored at 4 °C, and sampled four times at one month intervals. Two-dimensional gel electrophoresis and MALDI-TOF–TOF MS were performed to examine the protein changes during the postharvest storage period. Results showed that 74 proteins were differentially regulated, from which 56 proteins were identified by blasting against NCBInr (green plant) and EST_viridiplantae databases. All identified proteins were then classified into functional classes according to known biosynthetic pathways, including C-compound and carbohydrate metabolism, amino acid metabolism and response to storage environmental stimuli. In addition, subcellular location and time-dependent accumulation trends of differentially accumulated proteins associated with fruit quality were analyzed. To understand the relationships between these differentially accumulated proteins and fruit quality changes, composition of organic acids (malic acid, citric acid and quinic acid) and soluble sugars (fructose, glucose and sucrose) were investigated. Possible mechanisms responsible for fruit quality change in ponkan fruit during storage are discussed.

Amino acid deprivation induces CREBZF/Zhangfei expression via an AARE-like element in the promoter

CREBZF (also called ZF or Zhangfei) is a basic region–leucine zipper transcription factor that has been implicated in the herpesvirus infection cycle and related cellular processes. Since ATF4 is known to play a key role in cellular responses to various ER stresses as well as amino acid deprivation, we sought to examine the potential involvement of CREBZF in the amino acid response (AAR). We found that the CREBZF protein was induced by amino acid deprivation in the canine MDCK cells. We subsequently cloned a canine CREBZF promoter region (−1767 bp to +1 bp) that responds to amino acid limitation. Using deletion mapping and site-directed mutagenesis, we identified a 9-bp sequence 5′-ATTCACTCA-3′ in the promoter (−1227 to −1219), deletion of which resulted in a complete loss of inducibility by amino acid deprivation. This sequence is similar to the known amino acid response elements (AAREs) found in other AAR-inducible genes, such as CHOP (C/EBP homologous protein, also known as GADD153). These results suggest that CREBZF may be an amino acid stress sensor. Considering the AARE-like sequence found in CREBZF and other similarities between CREBZF and CHOP, we postulate that CREBZF and CHOP may be two sensors that regulate different yet related signaling pathways governing the AAR.

The Fate of Amino Acids During Simulated Meteoritic Impact

Delivery of prebiotic molecules, such as amino acids and peptides, in meteoritic/micrometeoritic materials to early Earth during the first 500 million years is considered to be one of the main processes by which the building blocks of life arrived on Earth. In this context, we present a study in which the effects of impact shock on amino acids and a peptide in artificial meteorites composed of saponite clay were investigated. The samples were subjected to pressures ranging from 12-28.9 GPa, which simulated impact velocities of 2.4-5.8 km/s for typical silicate-silicate impacts on Earth. Volatilization was determined by weight loss measurement, and the amino acid and peptide response was analyzed by gas chromatography-mass spectrometry. For all compounds, degradation increased with peak pressure. At the highest shock pressures, amino acids with an alkyl side chain were more resistant than those with functional side chains. The peptide cleaved into its two primary amino acids. Some chiral amino acids experienced partial racemization during the course of the experiment. Our data indicate that impact shock may act as a selective filter to the delivery of extraterrestrial amino acids via carbonaceous chondrites.

Identification of a Novel Amino Acid Response Pathway Triggering ATF2 Phosphorylation in Mammals

It has been well established that amino acid availability can control gene expression. Previous studies have shown that amino acid depletion induces transcription of the ATF3 (activation transcription factor 3) gene through an amino acid response element (AARE) located in its promoter. This event requires phosphorylation of activating transcription factor 2 (ATF2), a constitutive AARE-bound factor. To identify the signaling cascade leading to phosphorylation of ATF2 in response to amino acid starvation, we used an individual gene knockdown approach by small interfering RNA transfection. We identified the mitogen-activated protein kinase (MAPK) module MEKK1/MKK7/JNK2 as the pathway responsible for ATF2 phosphorylation on the threonine 69 (Thr69) and Thr71 residues. Then, we progressed backwards up the signal transduction pathway and showed that the GTPase Rac1/Cdc42 and the protein Galpha12 control the MAPK module, ATF2 phosphorylation, and AARE-dependent transcription. Taken together, our data reveal a new signaling pathway activated by amino acid starvation leading to ATF2 phosphorylation and subsequently positively affecting the transcription of amino acid-regulated genes.

A Bifunctional Intronic Element Regulates the Expression of the Arginine/Lysine Transporter Cat-1 via Mechanisms Involving the Purine-rich Element Binding Protein A (Purα)

Expression of the arginine/lysine transporter Cat-1 is highly induced in proliferating and stressed cells via mechanisms that include transcriptional activation. A bifunctional INE (intronic element) within the first intron of the Cat-1 gene was identified and characterized in this study. The INE had high sequence homology to an amino acid response element and was shown to act as a transcriptional enhancer in unstressed cells by binding the transcription factor, purine-rich element binding protein A (Pur alpha). During endoplasmic reticulum stress, binding of Pur alpha to the INE decreased; the element acted as a positive regulator in early stress by binding of the transcription factor ATF4 and as a negative regulator in prolonged stress by binding the stress-induced C/EBP family member, CHOP. We conclude that transcriptional control of the Cat-1 gene is tightly controlled by multiple cis-DNA elements, contributing to regulation of cationic amino acid transport for cell growth and proliferation. In addition, we propose that genes may use stress-response elements such as the INE to support basal expression in the absence of stress.

Distinctive Responses to Nitrogen Starvation in the Dominant Active Mutants of the Fission Yeast Rheb GTPase

Rheb, a Ras-like small GTPase conserved from human to yeast, controls Tor kinase and plays a central role in the regulation of cell growth depending on extracellular conditions. Rhb1 (a fission yeast homolog of Rheb) regulates amino acid uptake as well as response to nitrogen starvation. In this study, we generated two mutants, rhb1-DA4 and rhb1-DA8, and characterized them genetically. The V17A mutation within the G1 box defined for the Ras-like GTPases was responsible for rhb1-DA4 and Q52R I76F within the switch II domain for rhb1-DA8. In fission yeast, two events--the induction of the meiosis-initiating gene mei2+ and cell division without cell growth--are a typical response to nitrogen starvation. Under nitrogen-rich conditions, Rheb stimulates Tor kinase, which, in turn, suppresses the response to nitrogen starvation. While amino acid uptake was prevented by both rhb1-DA4 and rhb1-DA8 in a dominant fashion, the response to nitrogen starvation was prevented only by rhb1-DA4. rhb1-DA8 thereby allowed genetic dissection of the Rheb-dependent signaling cascade. We postulate that the signaling cascade may branch below Rhb1 or Tor2 and regulate the amino acid uptake and response to nitrogen starvation independently.

Cytokine-dependent and–independent gene expression changes and cell cycle block revealed in Trypanosoma cruzi-infected host cells by comparative mRNA profiling

BackgroundThe requirements for growth and survival of the intracellular pathogen Trypanosoma cruzi within mammalian host cells are poorly understood. Transcriptional profiling of the host cell response to infection serves as a rapid read-out for perturbation of host physiology that, in part, reflects adaptation to the infective process. Using Affymetrix oligonucleotide array analysis we identified common and disparate host cell responses triggered by T. cruzi infection of phenotypically diverse human cell types.ResultsWe report significant changes in transcript abundance in T. cruzi-infected fibroblasts, endothelial cells and smooth muscle cells (2852, 2155 and 531 genes respectively; fold-change ≥ 2, p-value < 0.01) 24 hours post-invasion. A prominent type I interferon response was observed in each cell type, reflecting a secondary response to secreted cytokine in infected cultures. To identify a core cytokine-independent response in T. cruzi-infected fibroblasts and endothelial cells transwell plates were used to distinguish cytokine-dependent and -independent gene expression profiles. This approach revealed the induction of metabolic and signaling pathways involved in cell proliferation, amino acid catabolism and response to wounding as common themes in T. cruzi-infected cells. In addition, the downregulation of genes involved in mitotic cell cycle and cell division predicted that T. cruzi infection may impede host cell cycle progression. The observation of impaired cytokinesis in T. cruzi-infected cells, following nuclear replication, confirmed this prediction.ConclusionMetabolic pathways and cellular processes were identified as significantly altered at the transcriptional level in response to T. cruzi infection in a cytokine-independent manner. Several of these alterations are supported by previous studies of T. cruzi metabolic requirements or effects on the host. However, our methods also revealed a T. cruzi-dependent block in the host cell cycle, at the level of cytokinesis, previously unrecognized for this pathogen-host cell interaction.

Changes in Basal and Insulin and Amino Acid Response of Whole Body and Skeletal Muscle Proteins in Obese Men

Obesity-related insulin resistance of glucose and lipid metabolism may also affect protein kinetics, notably at the muscle level. We hypothesized that muscle protein response to insulin and amino acid is blunted during obesity. Total (Tot) and mitochondrial (Mit) muscle proteins fractional synthesis rates (FSR) together with whole-body protein kinetics (WB) have been determined in postabsorptive state (PA) and during a hyperinsulinemic, hyperaminoacidemic, euglycemic clamp by using a continuous infusion of (13)C-leucine in six obese and eight nonobese subjects. Responses of WB glucose disposal rate and protein breakdown to insulin and amino acid infusion were significantly lower in obese than in nonobese subjects (P < 0.05). In PA, Tot and Mit FSR were significantly lower (P < 0.05) in obese (Tot, 0.044 +/- 0.005% . h(-1); Mit, 0.064 +/- 0.008% . h(-1)) in comparison with nonobese subjects (Tot, 0.082 +/- 0.010% . h(-1); Mit, 0.140 +/- 0.006% . h(-1)). Tot FSR was similarly stimulated by insulin and amino acid in both groups (0.094 +/- 0.013 vs. 0.117 +/- 0.006% . h(-1), obese vs. nonobese; P < 0.05). Mit FSR was increased in nonobese subjects (0.179 +/- 0.007% . h(-1); P < 0.05) but not in obese subjects (0.078 +/- 0.012% . h(-1); P = not significant). The obesity-related impairment of protein metabolism is characterized by 1) a reduced turnover rate of skeletal muscle proteins in PA; 2) a lack of stimulation of mitochondrial protein synthesis by insulin and amino acid; and 3) a lower inhibition of WB proteolysis by insulin and amino acid. Alterations of selective muscle protein kinetics may predispose obese subjects to muscle metabolic dysfunction leading to type 2 diabetes.

The Wnt pathway antagonist DKK1 is upregulated by amino acid deficiency in a human colon cancer cell line.

Amino acid response (AAR) pathway is activated when cells are deprived of amino acid. Activation of the AAR pathway causes upregulation of downstream genes. In the present study, we observed that the Wnt signaling pathway, playing an important role in development and tumorigenesis, is involved in this process. DKK1, an antagonist of the Wnt pathway, was significantly induced at mRNA level in the human colon cancer cell line SW480 after the removal of total amino acids and reached the peak of expression at 8hr. A key target of the AAR pathway, asparagine synthetase was also upregulated. Blocking the availability of a single amino acid histidine with histidinol also caused activation of the AAR pathway and led to an increase in DKK1 expression that is similar to total amino acid limitation. Blocking of the phosphorylation of ERk1/2 abolished the induction of DKK1. Analysis of mRNA stability showed that the induction was partly due to transcriptional regulation. Luciferase reporter assay was conducted to map the basal promoter and amino acid responsive element. Chromatin Immunoprecipitation assay was used to define transcription factor binding and chromatin modification induced by amino acid limitation. In conclusion, the present study showed that the Wnt signaling pathway is a potential downstream target of AAR pathway through the regulation of one of the antagonist, DKK1.Grant Funding SourceCIAF

Prenatal Low Protein Diet Influences Placental Gene Expression In Rats

Placental function and efficiency has been identified as a strong predictor of fetal growth and development. Because placenta acts as the only means for the fetus to obtain its nutrients, changes in placental morphology have dire consequences for fetal size and development. However, there is little data showing what physiological or external factors may cause these changes. This study intended to show that nutrition, specifically a low protein diet, contributes to altered expression of genes associated with placental health and transport capacity, including Igf2, H19, Atf3, Atf4, Dnmt1, Peg3, Glut1 and Snat2. Feeding pregnant rats a 9% low protein (LP) diet throughout gestation did not affect food intake or maternal weight. Atf3, but not Atf4 expression increased in LP placentas and there was a trend of increase in Igf2 and H19 expression. There was also a significant decrease in Peg3 and Dnmt1 expression in the LP group, and a significant increase in Glut1 and Snat2 expression. We did not detect changes in DNA methylation in many of these genes, indicating that control of gene expression for these genes is accomplished by means other than DNA methylation. Our study showed that mammalian amino acid response (AAR) pathway in placenta is up‐regulated by prenatal LP diet and this pathway can potentially cause other changes in placental gene expression.

Maternal Protein Restriction Alters Histone Modifications and Glucose Transporter 4 (GLUT4) Expression in Female Rat Skeletal Muscle during Postnatal Development

Maternal nutrition during pregnancy is an important intrauterine environmental factor associated with potential disease risk in later life. In this study, we investigated how epigenetic regulation of the muscle glucose transporter GLUT4 is modified in the offspring. Timed‐pregnant Sprague Dawley rats were fed with one of two isocaloric diets, control (18% casein) or low protein diet (9% casein) during gestation period. At birth, all mothers received a standardized meal ad libitum. The skeletal muscle was removed when pups were 38 days old. GLUT4 mRNA expression in female rat skeletal muscle was increased significantly by maternal low protein diet. An increase of expression in genes associated with glucose metabolism, such as CEBPβ and Nur77, was observed in female pups whose mothers were fed with low protein diet. Employing ChIP, we observed that modifications of histone acetylation and methylation at GLUT4 promoter region in female pup muscle were linked with the alteration in GLUT4 expression. However, the increased GLUT4 expression was not mediated by amino acid response pathway or DNA demethylation. These results suggest that a maternal diet limited in protein epigenetically alters fetal chromatin via covalent modifications of histones and subsequently alters gene‐specific expression.