Allosteric Regulation of a Protein Acetyltransferase in Micromonospora aurantiaca by the Amino Acids Cysteine and Arginine

Xu, Jun-Yu; You, Di; Leng, Pei-Qiang; Ye, Bang‐Ce

doi:10.1074/jbc.m114.579078

Cited by 21 publications

(46 citation statements)

References 39 publications

(40 reference statements)

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“…GCN5 is highly conserved across species (39) and was originally identified in yeast as an essential component for proper activation of the transcriptional response to amino acid starvation mediated by GCN4 (40). It was recently shown in lower organisms that amino acids can activate acetyltransferase activity of GCN5-related N-acetyltransferase (GNAT) protein (41). Whether GCN5 is a conserved component that can be activated in response to amino acid availability is of particular interest.…”

Section: Methionine Regulates Pgc-1␣ Activitymentioning

confidence: 99%

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

Tavares

Sharabi

Dominy

et al. 2016

Journal of Biological Chemistry

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Methionine is an essential sulfur amino acid that is engaged in key cellular functions such as protein synthesis and is a precursor for critical metabolites involved in maintaining cellular homeostasis. In mammals, in response to nutrient conditions, the liver plays a significant role in regulating methionine concentrations by altering its flux through the transmethylation, transsulfuration, and transamination metabolic pathways. A comprehensive understanding of how hepatic methionine metabolism intersects with other regulatory nutrient signaling and transcriptional events is, however, lacking. Here, we show that methionine and derived-sulfur metabolites in the transamination pathway activate the GCN5 acetyltransferase promoting acetylation of the transcriptional coactivator PGC-1␣ to control hepatic gluconeogenesis. Methionine was the only essential amino acid that rapidly induced PGC-1␣ acetylation through activating the GCN5 acetyltransferase. Experiments employing metabolic pathway intermediates revealed that methionine transamination, and not the transmethylation or transsulfuration pathways, contributed to methionine-induced PGC-1␣ acetylation. Moreover, aminooxyacetic acid, a transaminase inhibitor, was able to potently suppress PGC-1␣ acetylation stimulated by methionine, which was accompanied by predicted alterations in PGC-1␣-mediated gluconeogenic gene expression and glucose production in primary murine hepatocytes. Methionine administration in mice likewise induced hepatic PGC-1␣ acetylation, suppressed the gluconeogenic gene program, and lowered glycemia, indicating that a similar phenomenon occurs in vivo. These results highlight a communication between methionine metabolism and PGC-1␣-mediated hepatic gluconeogenesis, suggesting that influencing methionine metabolic flux has the potential to be therapeutically exploited for diabetes treatment.Amino acid sensing is a critical mechanism that cells employ to maintain homeostasis and enable survival upon fluctuations in the nutritional environment (1, 2). Despite variability in side chain chemical structure, cells are able to distinguish the distinct amino acids through the use of specific tRNAs. The amino acid protein sensor GCN2 detects deficiencies in amino acids in a tRNA-dependent manner and reprograms cellular metabolism to modulate energy consumption and maintain homeostasis (3, 4). Although mTORC1 is not considered to be a direct sensor of amino acid levels, it functions similarly to GCN2 to alter metabolic pathways in response to amino acid deprivation (5). Methionine, a dietary essential amino acid, is among the more toxic of the L-amino acids, and hence its abundance needs to be precisely regulated (6, 7). The liver is the primary organ concerned with the detoxification of methionine, and hence its ability to sense alterations in concentrations of this amino acid is vital (8). The existence of any cross-talk between hepatic methionine metabolism and other nutrient-regulated metabolic processes in the liver, such as glucose production, requi...

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Section: Methionine Regulates Pgc-1␣ Activitymentioning

confidence: 99%

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

Tavares

Sharabi

Dominy

et al. 2016

Journal of Biological Chemistry

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confidence: 99%

“…Following the discovery of acetylation of the Salmonella enterica acetyl coenzyme A (acetyl-CoA) synthetase in 2002 (9), this type of PTM has also emerged as an important metabolic regulatory mechanism in bacteria. In the last decade, lysine acetylation of proteins has also been reported in other organisms (6,7,(10)(11)(12)(13)(14)(15)(16).Protein lysine acetylation can occur via either enzymatic or nonenzymatic acetylation, such as chemical acetylation. Intracellular acetyl phosphate (AcP) plays a critical role in a chemical acetylation reaction; for example, AcP has been shown to chemically acetylate histones, serum albumin, and synthetic polylysine in vitro (17).…”

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confidence: 99%

“…Recently, we found that the amino acid-binding (ACT) domain is fused to the GNAT acetyltransferase in Micromonospora aurantiaca (MaKat), which confers amino acid-induced allosteric regulation (11). Similarly, fusion of the cAMP domain to the GNAT domain can occur in response to various stress conditions (i.e., cAMP concentration) to modulate the activity of a GNAT protein acetyltransferase in mycobacteria (27)(28)(29)(30).…”

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confidence: 99%

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Regulation of a Protein Acetyltransferase in Myxococcus xanthus by the Coenzyme NADP ⁺

Liu

2016

J Bacteriol

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NADP؉ is a vital cofactor involved in a wide variety of activities, such as redox potential and cell death. Here, we show that NADP ؉ negatively regulates an acetyltransferase from Myxococcus xanthus, Mxan_3215 (MxKat), at physiologic concentrations. MxKat possesses an NAD(P)-binding domain fused to the Gcn5-type N-acetyltransferase (GNAT) domain. We used isothermal titration calorimetry (ITC) and a coupled enzyme assay to show that NADP ؉ bound to MxKat and that the binding had strong effects on enzyme activity. The Gly11 residue of MxKat was confirmed to play an important role in NADP ؉ binding using sitedirected mutagenesis and circular dichroism spectrometry. In addition, using mass spectrometry, site-directed mutagenesis, and a coupling enzymatic assay, we demonstrated that MxKat acetylates acetyl coenzyme A (acetyl-CoA) synthetase (Mxan_2570) at Lys622 in response to changes in NADP ؉ concentration. Collectively, our results uncovered a mechanism of protein acetyltransferase regulation by the coenzyme NADP ؉ at physiological concentrations, suggesting a novel signaling pathway for the regulation of cellular protein acetylation. IMPORTANCE Microorganisms have developed various protein posttranslational modifications (PTMs), which enable cells to respond quickly to changes in the intracellular and extracellular milieus. This work provides the first biochemical characterization of a protein acetyltransferase (MxKat) that contains a fusion between a GNAT domain and NADP؉ -binding domain with Rossmann folds, and it demonstrates a novel signaling pathway for regulating cellular protein acetylation in M. xanthus. We found that NADP ؉ specifically binds to the Rossmann fold of MxKat and negatively regulates its acetyltransferase activity. This finding provides novel insight for connecting cellular metabolic status (NADP ؉ metabolism) with levels of protein acetylation, and it extends our understanding of the regulatory mechanisms underlying PTMs.T he dynamic and reversible mechanism of protein acetylation is an important regulatory posttranslational modification (PTM), which controls numerous cellular processes in the three kingdoms of life (1-4). Recent studies have identified Ͼ4,500 acetylated proteins, ranging from transcription factors and ribosomal proteins to many metabolic enzymes that are related to glycolysis, gluconeogenesis, the tricarboxylic acid (TCA) cycle, and fatty acid, nitrogen, and carbon metabolism (3,(5)(6)(7)(8). Following the discovery of acetylation of the Salmonella enterica acetyl coenzyme A (acetyl-CoA) synthetase in 2002 (9), this type of PTM has also emerged as an important metabolic regulatory mechanism in bacteria. In the last decade, lysine acetylation of proteins has also been reported in other organisms (6,7,(10)(11)(12)(13)(14)(15)(16).Protein lysine acetylation can occur via either enzymatic or nonenzymatic acetylation, such as chemical acetylation. Intracellular acetyl phosphate (AcP) plays a critical role in a chemical acetylation reaction; for example, AcP has been s...

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“…In Mycobacterium tuberculosis and Mycobacterium smegmatis, cAMP directly activates the protein acetyltransferases MtKat (Rv0998) and MsKat (MSMEG_5458) by binding to a cyclic nucleotide-binding domain that is fused to the N terminus of the catalytic GNAT domain (20). Recently, we found that an amino acid-binding domain (ACT domain) fused to the GNAT acetyltransferase of Micromonospora aurantica is used to exert amino acid-induced allosteric regulation of the enzyme (21). Thus, it is likely that the protein acetyltransferase enzymes are carefully regulated at the transcriptional and posttranslational levels in response to changes of the intracellular signals that control the Significance Nitrogen availability influences morphogenesis, antibiotic production, and virulence/pathogenicity in actinomycetes.…”

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Sirtuin-dependent reversible lysine acetylation of glutamine synthetases reveals an autofeedback loop in nitrogen metabolism

You

Yin

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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In cells of all domains of life, reversible lysine acetylation modulates the function of proteins involved in central cellular processes such as metabolism. In this study, we demonstrate that the nitrogen regulator GlnR of the actinomycete Saccharopolyspora erythraea directly regulates transcription of the acuA gene (SACE_5148), which encodes a Gcn5-type lysine acetyltransferase. We found that AcuA acetylates two glutamine synthetases (GlnA1 and GlnA4) and that this lysine acetylation inactivated GlnA4 (GSII) but had no significant effect on GlnA1 (GSI-β) activity under the conditions tested. Instead, acetylation of GlnA1 led to a gain-of-function that modulated its interaction with the GlnR regulator and enhanced GlnR–DNA binding. It was observed that this regulatory function of acetylated GSI-β enzymes is highly conserved across actinomycetes. In turn, GlnR controls the catalytic and regulatory activities (intracellular acetylation levels) of glutamine synthetases at the transcriptional and posttranslational levels, indicating an autofeedback loop that regulates nitrogen metabolism in response to environmental change. Thus, this GlnR-mediated acetylation pathway provides a signaling cascade that acts from nutrient sensing to acetylation of proteins to feedback regulation. This work presents significant new insights at the molecular level into the mechanisms underlying the regulation of protein acetylation and nitrogen metabolism in actinomycetes.

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Allosteric Regulation of a Protein Acetyltransferase in Micromonospora aurantiaca by the Amino Acids Cysteine and Arginine

Cited by 21 publications

References 39 publications

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

Regulation of a Protein Acetyltransferase in Myxococcus xanthus by the Coenzyme NADP ⁺

Sirtuin-dependent reversible lysine acetylation of glutamine synthetases reveals an autofeedback loop in nitrogen metabolism

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Allosteric Regulation of a Protein Acetyltransferase in Micromonospora aurantiaca by the Amino Acids Cysteine and Arginine

Cited by 21 publications

References 39 publications

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

The Methionine Transamination Pathway Controls Hepatic Glucose Metabolism through Regulation of the GCN5 Acetyltransferase and the PGC-1α Transcriptional Coactivator

Regulation of a Protein Acetyltransferase in Myxococcus xanthus by the Coenzyme NADP +

Sirtuin-dependent reversible lysine acetylation of glutamine synthetases reveals an autofeedback loop in nitrogen metabolism

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Regulation of a Protein Acetyltransferase in Myxococcus xanthus by the Coenzyme NADP ⁺