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MSystems Dec 2022Antibiotic resistance is increasingly becoming a challenge to public health. The regulation of bacterial metabolism by post-translational modifications (PTMs) has been...
Antibiotic resistance is increasingly becoming a challenge to public health. The regulation of bacterial metabolism by post-translational modifications (PTMs) has been widely studied. However, the mechanism underlying the regulation of acetylation in bacterial resistance to antibiotics is still unknown. Here, we performed a quantitative analysis of the acetylated proteome of a wild-type (WT) Escherichia coli (E. coli) sensitive strain and ampicillin- (Re-Amp), kanamycin- (Re-Kan), and polymyxin B-resistant (Re-Pol) strains. Based on bioinformatics analysis combined with biochemical validations, we found a common regulatory mechanism between the different resistant strains. Our results showed that protein acetylation negatively regulates bacterial metabolism to regulate antibiotic resistance and positively regulates bacterial motility. Further analyses revealed that key enzymes in various metabolic pathways were differentially acetylated. In particular, pyruvate kinase (PykF), a glycolytic enzyme that regulates bacterial metabolism, and its acetylated form were highly expressed in the three resistant strains and were identified as reversibly acetylated by the deacetylase CobB and the acetyl-transferase PatZ (peptidyl-lysine -acetyltransferase). Results showed that PykF also could be acetylated by nonenzymatic acetyl phosphatase (AcP) . Furthermore, the deacetylation of Lys413 in PykF increased PykF enzymatic activity by changing the conformation of its ATP binding site, resulting in an increase in energy production which, in turn, increased the sensitivity of drug-resistant strains to antibiotics. This study provides novel insights for understanding bacterial resistance and lays the foundation for future research on the regulation of acetylation in antibiotic-resistant strains. The misuse of antibiotics has resulted in the emergence of many antibiotic-resistant strains which seriously threaten human health. Protein post-translational modifications, especially acetylation, tightly control bacterial metabolism. However, the comprehensive mechanism underlying the regulation of acetylation in bacterial resistance remains unexplored. Here, acetylation was found to positively regulate bacterial motility and negatively regulate energy metabolism, which was common in all antibiotic-resistant strains. Moreover, the acetylation and deacetylation process of PykF was uncovered, and deacetylation of the Lys 413 in PykF was found to contribute to bacterial sensitivity to antibiotics. This study provides a new direction for research on the development of bacterial resistance through post-translational modifications and a theoretical basis for developing antibacterial drugs.
Topics: Humans; Escherichia coli; Lysine; Acetylation; Protein Processing, Post-Translational; Anti-Bacterial Agents; Lysine Acetyltransferases; Pyruvate Kinase; Drug Resistance, Microbial
PubMed: 36286553
DOI: 10.1128/msystems.00649-22 -
Nature Communications Sep 2022Covalent attachment of ubiquitin (Ub) to proteins is a highly versatile posttranslational modification. Moreover, Ub is not only a modifier but itself is modified by...
Covalent attachment of ubiquitin (Ub) to proteins is a highly versatile posttranslational modification. Moreover, Ub is not only a modifier but itself is modified by phosphorylation and lysine acetylation. However, the functional consequences of Ub acetylation are poorly understood. By generation and comprehensive characterization of all seven possible mono-acetylated Ub variants, we show that each acetylation site has a particular impact on Ub structure. This is reflected in selective usage of the acetylated variants by different E3 ligases and overlapping but distinct interactomes, linking different acetylated variants to different cellular pathways. Notably, not only electrostatic but also steric effects contribute to acetylation-induced changes in Ub structure and, thus, function. Finally, we provide evidence that p300 acts as a position-specific Ub acetyltransferase and HDAC6 as a general Ub deacetylase. Our findings provide intimate insights into the structural and functional consequences of Ub acetylation and highlight the general importance of Ub acetylation.
Topics: Acetylation; Acetyltransferases; Lysine; Protein Processing, Post-Translational; Static Electricity; Ubiquitin; Ubiquitin-Protein Ligases
PubMed: 36114200
DOI: 10.1038/s41467-022-33087-1 -
Cells Mar 2020-acetylation of sialic acid residues is one of the main modifications of gangliosides, and modulates ganglioside functions. -acetylation of gangliosides is dependent on... (Review)
Review
-acetylation of sialic acid residues is one of the main modifications of gangliosides, and modulates ganglioside functions. -acetylation of gangliosides is dependent on sialyl--acetyltransferases and sialyl--acetyl-esterase activities. CAS1 Domain-Containing Protein 1 (CASD1) is the only human sialyl--acetyltransferases (SOAT) described until now. -acetylated ganglioside species are mainly expressed during embryonic development and in the central nervous system in healthy adults, but are re-expressed during cancer development and are considered as markers of cancers of neuroectodermal origin. However, the specific biological roles of -acetylated gangliosides in developing and malignant tissues have not been extensively studied, mostly because of the requirement of specific approaches and tools for sample preparation and analysis. In this review, we summarize our current knowledge of ganglioside biosynthesis and expression in normal and pathological conditions, of ganglioside -acetylation analysis and expression in cancers, and of the possible use of -acetylated gangliosides as targets for cancer immunotherapy.
Topics: Acetylation; Animals; Gangliosides; Humans; Immunotherapy; Molecular Targeted Therapy; N-Acetylneuraminic Acid; Neoplasms
PubMed: 32192217
DOI: 10.3390/cells9030741 -
Experimental & Molecular Medicine Jul 2018Post-translational modifications (PTMs) are chemical alterations that occur in proteins that play critical roles in various cellular functions. Lysine acetylation is an... (Review)
Review
Post-translational modifications (PTMs) are chemical alterations that occur in proteins that play critical roles in various cellular functions. Lysine acetylation is an important PTM in eukaryotes, and it is catalyzed by lysine acetyltransferases (KATs). KATs transfer acetyl-coenzyme A to the internal lysine residue of substrate proteins. Arrest defective 1 (ARD1) is a member of the KAT family. Since the identification of its KAT activity 15 years ago, many studies have revealed that diverse cellular proteins are acetylated by ARD1. ARD1-mediated lysine acetylation is a key switch that regulates the enzymatic activities and biological functions of proteins and influences cell biology from development to pathology. In this review, we summarize protein lysine acetylation mediated by ARD1 and describe the biological meanings of this modification.
Topics: Acetylation; Animals; Humans; Lysine; N-Terminal Acetyltransferase A; N-Terminal Acetyltransferase E; Protein Processing, Post-Translational
PubMed: 30054464
DOI: 10.1038/s12276-018-0100-7 -
Genes & Development Apr 2018Fluctuations in acetyl-coenzyme A (acetyl-CoA) levels have been previously associated with changes in global histone acetylation and gene expression. The study by Lee... (Review)
Review
Fluctuations in acetyl-coenzyme A (acetyl-CoA) levels have been previously associated with changes in global histone acetylation and gene expression. The study by Lee and colleagues (pp. 497-511) in this issue of demonstrates that acetyl-CoA can promote the up-regulation of cell migration- and adhesion-related genes in glioblastoma by controlling Ca-NFAT (nuclear factor of activated T cells) signaling.
Topics: Acetyl Coenzyme A; Acetylation; Adult; Calcium; Cell Adhesion; Glioblastoma; Histones; Humans; Transcription, Genetic
PubMed: 29692354
DOI: 10.1101/gad.315168.118 -
Journal of Bacteriology Aug 2017-Lysine acetylation is now recognized as an abundant posttranslational modification (PTM) that influences many essential biological pathways. Advancements in mass... (Review)
Review
-Lysine acetylation is now recognized as an abundant posttranslational modification (PTM) that influences many essential biological pathways. Advancements in mass spectrometry-based proteomics have led to the discovery that bacteria contain hundreds of acetylated proteins, contrary to the prior notion of acetylation events being rare in bacteria. Although the mechanisms that regulate protein acetylation are still not fully defined, it is understood that this modification is finely tuned via both enzymatic and nonenzymatic mechanisms. The opposing actions of Gcn5-related -acetyltransferases (GNATs) and deacetylases, including sirtuins, provide the enzymatic control of lysine acetylation. A nonenzymatic mechanism of acetylation has also been demonstrated and proven to be prominent in bacteria, as well as in mitochondria. The functional consequences of the vast majority of the identified acetylation sites remain unknown. From studies in mammalian systems, acetylation of critical lysine residues was shown to impact protein function by altering its structure, subcellular localization, and interactions. It is becoming apparent that the same diversity of functions can be found in bacteria. Here, we review current knowledge of the mechanisms and the functional consequences of acetylation in bacteria. Additionally, we discuss the methods available for detecting acetylation sites, including quantitative mass spectrometry-based methods, which promise to promote this field of research. We conclude with possible future directions and broader implications of the study of protein acetylation in bacteria.
Topics: Acetylation; Bacteria; Bacterial Proteins; Mass Spectrometry; Protein Processing, Post-Translational; Proteome
PubMed: 28439035
DOI: 10.1128/JB.00107-17 -
The Journal of Biological Chemistry Aug 2021Sialic acids are nine-carbon sugars that frequently cap glycans at the cell surface in cells of vertebrates as well as cells of certain types of invertebrates and... (Review)
Review
Sialic acids are nine-carbon sugars that frequently cap glycans at the cell surface in cells of vertebrates as well as cells of certain types of invertebrates and bacteria. The nine-carbon backbone of sialic acids can undergo extensive enzymatic modification in nature and O-acetylation at the C-4/7/8/9 position in particular is widely observed. In recent years, the detection and analysis of O-acetylated sialic acids have advanced, and sialic acid-specific O-acetyltransferases (SOATs) and O-acetylesterases (SIAEs) that add and remove O-acetyl groups, respectively, have been identified and characterized in mammalian cells, invertebrates, bacteria, and viruses. These advances now allow us to draw a more complete picture of the biosynthetic pathway of the diverse O-acetylated sialic acids to drive the generation of genetically and biochemically engineered model cell lines and organisms with altered expression of O-acetylated sialic acids for dissection of their roles in glycoprotein stability, development, and immune recognition, as well as discovery of novel functions. Furthermore, a growing number of studies associate sialic acid O-acetylation with cancer, autoimmunity, and infection, providing rationale for the development of selective probes and inhibitors of SOATs and SIAEs. Here, we discuss the current insights into the biosynthesis and biological functions of O-acetylated sialic acids and review the evidence linking this modification to disease. Furthermore, we discuss emerging strategies for the design, synthesis, and potential application of unnatural O-acetylated sialic acids and inhibitors of SOATs and SIAEs that may enable therapeutic targeting of this versatile sialic acid modification.
Topics: Acetylation; Acetyltransferases; Animals; Biosynthetic Pathways; Carboxylic Ester Hydrolases; Disease; Glycoproteins; Humans; N-Acetylneuraminic Acid; Polysaccharides
PubMed: 34157283
DOI: 10.1016/j.jbc.2021.100906 -
Environmental Health Perspectives Mar 1983N-Substituted aromatic compounds can be metabolized in most species to N-acetylated derivatives that are themselves subject to further enzymatic transformations,... (Review)
Review
N-Substituted aromatic compounds can be metabolized in most species to N-acetylated derivatives that are themselves subject to further enzymatic transformations, including hydrolysis and N,O-acyltransfer. These proceses can either potentiate or ameliorate the biological responses to these N-substituted derivatives. Decreasing the levels of metabolites, such as arylhydroxylamines may, in some systems, reduce the probability of eliciting adverse biological effects. In others, arylhydroxamic acids produced by the acetylation of arylhydroxylamines may increase their potential for metabolic activation by N,O-acyltransfer. In the rabbit, rat and perhaps other species, the acetyl CoA-dependent N-acetyltransferase is also capable of activating arylhydroxamic acids by N-O-acyltransfer. These cytosolic organotriphosphate ester-resistant enzymes can utilize arylhydroxamic acid as a donor of the acetyl moiety in the acetyl transferase reaction and apparently are capable of activating arylhydroxamic acids because of their ability to O-acetylate the arylhydroxlamine. In mice, N-acetyltransferase and N,O-acetyltransferase seem not to exhibit this relationship. Enzymes from the microsomes of a number of species are also capable of activating arylhydroxamic acids. The particulate-bound enzymes are organotriphosphate ester-sensitive deacylases that are unable to form nucleic acid adducts on incubation with N-methoxy-N-acetylaminoarenes, substrates that are not capable of activation by N,O-acyltransfer. Thus, depending on the specificity of the enzymes involved, N-substituted aromatic compounds may be activated by N,O-acyltransfer during both the acetylation and deacylation process. The influence of this activation in the carcinogenic process is the object of continuing investigation.
Topics: Acetyl-CoA C-Acetyltransferase; Acetylation; Acetyltransferases; Acyltransferases; Amides; Animals; Carcinogens; Cytosol; Humans; Hydrolysis; Hydroxamic Acids; Microsomes; Polymorphism, Genetic; Species Specificity
PubMed: 6131820
DOI: 10.1289/ehp.834943 -
International Journal of Molecular... Nov 2020Elp3, the catalytic subunit of the eukaryotic Elongator complex, is a lysine acetyltransferase that acetylates the C5 position of wobble-base uridines (U) in transfer... (Review)
Review
Elp3, the catalytic subunit of the eukaryotic Elongator complex, is a lysine acetyltransferase that acetylates the C5 position of wobble-base uridines (U) in transfer RNAs (tRNAs). This Elongator-dependent RNA acetylation of anticodon bases affects the ribosomal translation elongation rates and directly links acetyl-CoA metabolism to both protein synthesis rates and the proteome integrity. Of note, several human diseases, including various cancers and neurodegenerative disorders, correlate with the dysregulation of Elongator's tRNA modification activity. In this review, we focus on recent findings regarding the structure of Elp3 and the role of acetyl-CoA during its unique modification reaction.
Topics: Acetylation; Animals; Base Sequence; Binding Sites; Histone Acetyltransferases; Humans; Lysine; Nerve Tissue Proteins; Peptide Chain Elongation, Translational; RNA Processing, Post-Transcriptional; RNA, Transfer; Uridine
PubMed: 33152999
DOI: 10.3390/ijms21218209 -
Advances in Experimental Medicine and... 2019Mitochondria have a central role in cellular metabolism and reversible post-translational modifications regulate activity of mitochondrial proteins. Thanks to advances...
Mitochondria have a central role in cellular metabolism and reversible post-translational modifications regulate activity of mitochondrial proteins. Thanks to advances in proteomics, lysine acetylation has arisen as an important post-translational modification in the mitochondrion. During acetylation an acetyl group is covalently attached to the epsilon amino group in the side chain of lysine residues using acetyl-CoA as the substrate donor. Therefore the positive charge is neutralized, and this can affect the function of proteins thereby regulating enzyme activity, protein interactions, and protein stability. The major deacetylase in mitochondria is SIRT3 whose activity regulates many mitochondrial enzymes. The method of choice for the analysis of acetylated proteins foresees the combination of mass spectrometry-based proteomics with affinity enrichment techniques. Beyond the identification of lysine-acetylated proteins, many studies are moving towards the characterization of acetylated patterns in different diseases. Indeed, modifications in lysine acetylation status can directly alter mitochondrial function and, therefore, be linked to human diseases such as metabolic diseases, cancer, myocardial injury and neurodegenerative diseases. Despite the progress in the characterization of different lysine acetylation sites, additional studies are needed to differentiate the specific changes with a significant biological relevance.
Topics: Acetylation; Humans; Lysine; Mitochondria; Mitochondrial Proteins; Phenotype; Protein Processing, Post-Translational
PubMed: 31452135
DOI: 10.1007/978-981-13-8367-0_4