ATP-Citrate Lyase Is Required for Production of Cytosolic Acetyl Coenzyme A and Development in Aspergillus nidulans

ABSTRACT Acetyl coenzyme A (CoA) is a central metabolite in carbon and energy metabolism and in the biosynthesis of cellular molecules. A source of cytoplasmic acetyl-CoA is essential for the production of fatty acids and sterols and for protein acetylation, including histone acetylation in the nucleus. In Saccharomyces cerevisiae and Candida albicans acetyl-CoA is produced from acetate by cytoplasmic acetyl-CoA synthetase, while in plants and animals acetyl-CoA is derived from citrate via ATP-citrate lyase. In the filamentous ascomycete Aspergillus nidulans, tandem divergently transcribed genes (aclA and aclB) encode the subunits of ATP-citrate lyase, and we have deleted these genes. Growth is greatly diminished on carbon sources that do not result in cytoplasmic acetyl-CoA, such as glucose and proline, while growth is not affected on carbon sources that result in the production of cytoplasmic acetyl-CoA, such as acetate and ethanol. Addition of acetate restores growth on glucose or proline, and this is dependent on facA, which encodes cytoplasmic acetyl-CoA synthetase, but not on the regulatory gene facB. Transcription of aclA and aclB is repressed by growth on acetate or ethanol. Loss of ATP-citrate lyase results in severe developmental effects, with the production of asexual spores (conidia) being greatly reduced and a complete absence of sexual development. This is in contrast to Sordaria macrospora, in which fruiting body formation is initiated but maturation is defective in an ATP-citrate lyase mutant. Addition of acetate does not repair these defects, indicating a specific requirement for high levels of cytoplasmic acetyl-CoA during differentiation. Complementation in heterokaryons between aclA and aclB deletions for all phenotypes indicates that the tandem gene arrangement is not essential.

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Functional Analyses of Two Acetyl Coenzyme A Synthetases in the Ascomycete Gibberella zeae

Eukaryotic Cell ◽

10.1128/ec.05071-11 ◽

2011 ◽

Vol 10 (8) ◽

pp. 1043-1052 ◽

Cited By ~ 44

Author(s):

Seunghoon Lee ◽

Hokyoung Son ◽

Jungkwan Lee ◽

Kyunghun Min ◽

Gyung Ja Choi ◽

...

Keyword(s):

Sexual Development ◽

Coenzyme A ◽

Lipid Synthesis ◽

Gibberella Zeae ◽

Atp Citrate Lyase ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Temporal Precision ◽

Acetyl Coenzyme

ABSTRACTAcetyl coenzyme A (acetyl-CoA) is a crucial metabolite for energy metabolism and biosynthetic pathways and is produced in various cellular compartments with spatial and temporal precision. Our previous study on ATP citrate lyase (ACL) inGibberella zeaerevealed that ACL-dependent acetyl-CoA production is important for histone acetylation, especially in sexual development, but is not involved in lipid synthesis. In this study, we deleted additional acetyl-CoA synthetic genes, the acetyl-CoA synthetases (ACSgenesACS1andACS2), to identify alternative acetyl-CoA production mechanisms for ACL. TheACS1deletion resulted in a defect in sexual development that was mainly due to a reduction in 1-palmitoyl-2-oleoyl-3-linoleoyl-rac-glycerol production, which is required for perithecium development and maturation. Another ACS coding gene,ACS2, has accessorial functions forACS1and has compensatory functions forACLas a nuclear acetyl-CoA producer. This study showed that acetate is readily generated during the entire life cycle ofG. zeaeand has a pivotal role in fungal metabolism. Because ACSs are components of the pyruvate-acetaldehyde-acetate pathway, this fermentation process might have crucial roles in various physiological processes for filamentous fungi.

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Genome Sequence of Serratia marcescens MSU97, a Plant-Associated Bacterium That Makes Multiple Antibiotics

Genome Announcements ◽

10.1128/genomea.01752-16 ◽

2017 ◽

Vol 5 (9) ◽

Cited By ~ 5

Author(s):

Miguel A. Matilla ◽

Zulema Udaondo ◽

Tino Krell ◽

George P. C. Salmond

Keyword(s):

Serratia Marcescens ◽

Genome Sequence ◽

Coenzyme A ◽

Acetyl Coa Carboxylase ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Acetyl Coenzyme ◽

Multiple Antibiotics

ABSTRACT Serratia marcescens MSU97 was isolated from the Guayana region of Venezuela due to its ability to suppress plant-pathogenic oomycetes. Here, we report the genome sequence of MSU97, which produces various antibiotics, including the bacterial acetyl-coenzyme A (acetyl-CoA) carboxylase inhibitor andrimid, the chlorinated macrolide oocydin A, and the red linear tripyrrole antibiotic prodigiosin.

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A Pathway for Degradation of Uracil to Acetyl Coenzyme A in Bacillus megaterium

Applied and Environmental Microbiology ◽

10.1128/aem.02837-19 ◽

2020 ◽

Vol 86 (7) ◽

Cited By ~ 2

Author(s):

Di Zhu ◽

Yifeng Wei ◽

Jinyu Yin ◽

Dazhi Liu ◽

Ee Lui Ang ◽

...

Keyword(s):

Energy Source ◽

Bacillus Megaterium ◽

Coenzyme A ◽

Catabolic Pathway ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Biochemical Pathways ◽

Acetyl Coenzyme

ABSTRACT Bacteria utilize diverse biochemical pathways for the degradation of the pyrimidine ring. The function of the pathways studied to date has been the release of nitrogen for assimilation. The most widespread of these pathways is the reductive pyrimidine catabolic pathway, which converts uracil into ammonia, carbon dioxide, and β-alanine. Here, we report the characterization of a β-alanine:pyruvate aminotransferase (PydD2) and an NAD+-dependent malonic semialdehyde dehydrogenase (MSDH) from a reductive pyrimidine catabolism gene cluster in Bacillus megaterium. Together, these enzymes convert β-alanine into acetyl coenzyme A (acetyl-CoA), a key intermediate in carbon and energy metabolism. We demonstrate the growth of B. megaterium in defined medium with uracil as its sole carbon and energy source. Homologs of PydD2 and MSDH are found in association with reductive pyrimidine pathway genes in many Gram-positive bacteria in the order Bacillales. Our study provides a basis for further investigations of the utilization of pyrimidines as a carbon and energy source by bacteria. IMPORTANCE Pyrimidine has wide occurrence in natural environments, where bacteria use it as a nitrogen and carbon source for growth. Detailed biochemical pathways have been investigated with focus mainly on nitrogen assimilation in the past decades. Here, we report the discovery and characterization of two important enzymes, PydD2 and MSDH, which constitute an extension for the reductive pyrimidine catabolic pathway. These two enzymes, prevalent in Bacillales based on our bioinformatics studies, allow stepwise conversion of β-alanine, a previous “end product” of the reductive pyrimidine degradation pathway, to acetyl-CoA as carbon and energy source.

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Conversion of 4-Hydroxybutyrate to Acetyl Coenzyme A and Its Anapleurosis in the Metallosphaera sedula 3-Hydroxypropionate/4-Hydroxybutyrate Carbon Fixation Pathway

Applied and Environmental Microbiology ◽

10.1128/aem.04146-13 ◽

2014 ◽

Vol 80 (8) ◽

pp. 2536-2545 ◽

Cited By ~ 17

Author(s):

Aaron B. Hawkins ◽

Michael W. W. Adams ◽

Robert M. Kelly

Keyword(s):

Carbon Fixation ◽

Coenzyme A ◽

Flux Analysis ◽

Central Metabolism ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Metallosphaera Sedula ◽

Acetyl Coenzyme ◽

Coa Ligase

ABSTRACTThe extremely thermoacidophilic archaeonMetallosphaera sedula(optimum growth temperature, 73°C, pH 2.0) grows chemolithoautotrophically on metal sulfides or molecular hydrogen by employing the 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) carbon fixation cycle. This cycle adds two CO2molecules to acetyl coenzyme A (acetyl-CoA) to generate 4HB, which is then rearranged and cleaved to form two acetyl-CoA molecules. Previous metabolic flux analysis showed that two-thirds of central carbon precursor molecules are derived from succinyl-CoA, which is oxidized to malate and oxaloacetate. The remaining one-third is apparently derived from acetyl-CoA. As such, the steps beyond succinyl-CoA are essential for completing the carbon fixation cycle and for anapleurosis of acetyl-CoA. Here, the final four enzymes of the 3HP/4HB cycle, 4-hydroxybutyrate-CoA ligase (AMP forming) (Msed_0406), 4-hydroxybutyryl-CoA dehydratase (Msed_1321), crotonyl-CoA hydratase/(S)-3-hydroxybutyryl-CoA dehydrogenase (Msed_0399), and acetoacetyl-CoA β-ketothiolase (Msed_0656), were produced recombinantly inEscherichia coli, combinedin vitro, and shown to convert 4HB to acetyl-CoA. Metabolic pathways connecting CO2fixation and central metabolism were examined using a gas-intensive bioreactor system in whichM. sedulawas grown under autotrophic (CO2-limited) and heterotrophic conditions. Transcriptomic analysis revealed the importance of the 3HP/4HB pathway in supplying acetyl-CoA to anabolic pathways generating intermediates inM. sedulametabolism. The results indicated that flux between the succinate and acetyl-CoA branches in the 3HP/4HB pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase (Pat)/Sir2-dependent system. Taken together, this work confirms the final four steps of the 3HP/4HB pathway, thereby providing the framework for examining connections between CO2fixation and central metabolism inM. sedula.

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Molecular Characterization of a Heteromeric ATP-Citrate Lyase That Generates Cytosolic Acetyl-Coenzyme A in Arabidopsis

PLANT PHYSIOLOGY ◽

10.1104/pp.008110 ◽

2002 ◽

Vol 130 (2) ◽

pp. 740-756 ◽

Cited By ~ 117

Author(s):

Beth L. Fatland ◽

Jinshan Ke ◽

Marc D. Anderson ◽

Wieslawa I. Mentzen ◽

Li Wei Cui ◽

...

Keyword(s):

Molecular Characterization ◽

Coenzyme A ◽

Atp Citrate Lyase ◽

Acetyl Coenzyme A ◽

Acetyl Coenzyme ◽

Citrate Lyase

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Role of Carnitine Acetyltransferases in Acetyl Coenzyme A Metabolism in Aspergillus nidulans

Eukaryotic Cell ◽

10.1128/ec.00295-10 ◽

2011 ◽

Vol 10 (4) ◽

pp. 547-555 ◽

Cited By ~ 18

Author(s):

Michael J. Hynes ◽

Sandra L. Murray ◽

Alex Andrianopoulos ◽

Meryl A. Davis

Keyword(s):

Fatty Acids ◽

Aspergillus Nidulans ◽

Intracellular Transport ◽

Coenzyme A ◽

Tricarboxylic Acid Cycle ◽

Essential Feature ◽

Malate Synthase ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Acetyl Coenzyme

ABSTRACTThe flow of carbon metabolites between cellular compartments is an essential feature of fungal metabolism. During growth on ethanol, acetate, or fatty acids, acetyl units must enter the mitochondrion for metabolism via the tricarboxylic acid cycle, and acetyl coenzyme A (acetyl-CoA) in the cytoplasm is essential for the biosynthetic reactions and for protein acetylation. Acetyl-CoA is produced in the cytoplasm by acetyl-CoA synthetase during growth on acetate and ethanol while β-oxidation of fatty acids generates acetyl-CoA in peroxisomes. The acetyl-carnitine shuttle in which acetyl-CoA is reversibly converted to acetyl-carnitine by carnitine acetyltransferase (CAT) enzymes is important for intracellular transport of acetyl units. In the filamentous ascomyceteAspergillus nidulans, a cytoplasmic CAT, encoded byfacC, is essential for growth on sources of cytoplasmic acetyl-CoA while a second CAT, encoded by theacuJgene, is essential for growth on fatty acids as well as acetate. We have shown that AcuJ contains an N-terminal mitochondrial targeting sequence and a C-terminal peroxisomal targeting sequence (PTS) and is localized to both peroxisomes and mitochondria, independent of the carbon source. Mislocalization of AcuJ to the cytoplasm does not result in loss of growth on acetate but prevents growth on fatty acids. Therefore, while mitochondrial AcuJ is essential for the transfer of acetyl units to mitochondria, peroxisomal localization is required only for transfer from peroxisomes to mitochondria. Peroxisomal AcuJ was not required for the import of acetyl-CoA into peroxisomes for conversion to malate by malate synthase (MLS), and export of acetyl-CoA from peroxisomes to the cytoplasm was found to be independent of FacC when MLS was mislocalized to the cytoplasm.

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Synthetic Biology for Engineering Acetyl Coenzyme A Metabolism in Yeast

mBio ◽

10.1128/mbio.02153-14 ◽

2014 ◽

Vol 5 (6) ◽

Cited By ~ 47

Author(s):

Jens Nielsen

Keyword(s):

Coenzyme A ◽

Pyruvate Dehydrogenase Complex ◽

High Yield ◽

Cell Factory ◽

Efficient Production ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Content Type ◽

Acetyl Coenzyme ◽

High Yields

ABSTRACT The yeast Saccharomyces cerevisiae is a widely used cell factory for the production of fuels, chemicals, and pharmaceuticals. The use of this cell factory for cost-efficient production of novel fuels and chemicals requires high yields and low by-product production. Many industrially interesting chemicals are biosynthesized from acetyl coenzyme A (acetyl-CoA), which serves as a central precursor metabolite in yeast. To ensure high yields in production of these chemicals, it is necessary to engineer the central carbon metabolism so that ethanol production is minimized (or eliminated) and acetyl-CoA can be formed from glucose in high yield. Here the perspective of generating yeast platform strains that have such properties is discussed in the context of a major breakthrough with expression of a functional pyruvate dehydrogenase complex in the cytosol.

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PanM, an Acetyl-Coenzyme A Sensor Required for Maturation of l-Aspartate Decarboxylase (PanD)

mBio ◽

10.1128/mbio.00158-12 ◽

2012 ◽

Vol 3 (4) ◽

Cited By ~ 8

Author(s):

Tara N. Stuecker ◽

Alex C. Tucker ◽

Jorge C. Escalante-Semerena

Keyword(s):

Coenzyme A ◽

Lysine Acetylation ◽

Acetyl Coa ◽

Transfer Event ◽

Acetyl Coenzyme A ◽

Content Type ◽

Acetyltransferase Activity ◽

Acetyl Coenzyme

ABSTRACTCoenzyme A (CoA) is essential for cellular chemistry in all forms of life. The pantothenate moiety of CoA is generated from the condensation of pantoate and β-alanine. β-Alanine is formed by decarboxylation ofl-aspartate catalyzed by PanD, a pyruvoyl enzyme that is synthesized by the cell as an inactive precursor (pro-PanD). Maturation of pro-PanD into PanD occurs via a self-cleavage event at residue Ser25, which forms the catalytic pyruvoyl moiety. We recently reported thatSalmonella entericaPanM was necessary for pro-PanD maturation, bothin vitroandin vivo. Notably, PanM is annotated as a Gcn5-likeN-acetyltransferase (GNAT), which suggested that lysine acetylation might be part of the mechanism of maturation. Here we show that PanM lacks acetyltransferase activity and that acetyl-CoA stimulates its activity. Results of experiments with nonhydrolyzable ethyl-CoA and genetically encoded acetyl-lysine-containing PanD support the conclusion that PanM-dependent pro-PanD maturation does not involve an acetyl transfer event. We also show that CoA binding to PanM is needed forin vivoactivity and that disruption of CoA binding prevents PanM from interacting with PanD. We conclude that PanM is a GNAT homologue that lost its acetyltransferase activity and evolved a new function as an acetyl-CoA sensor that can trigger the maturation of pro-PanD.IMPORTANCENε-lysine acetylation is increasingly being recognized as a widespread and important form of posttranslational regulation in bacteria. The acetyltransferases that catalyze these reactions are poorly characterized in bacteria. Based on annotation, most bacterial genomes contain several acetyltransferases, but the physiological roles of only a handful have been determined. Notably, a subset of putative acetyltransferases lack residues that are critical for activity in most biochemically characterized acetyltransferases. We show that one such putative acetyltransferase, PanM (formerly YhhK), lacks acetyltransferase activity but functions instead as an acetyl-coenzyme A (CoA) sensor. This work establishes the possibility that, like PanM, other putative acetyltransferases may have evolved new functions while retaining the ability to sense acetyl-CoA.

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Role of Acetyl Coenzyme A Synthesis and Breakdown in Alternative Carbon Source Utilization in Candida albicans

Eukaryotic Cell ◽

10.1128/ec.00253-08 ◽

2008 ◽

Vol 7 (10) ◽

pp. 1733-1741 ◽

Cited By ~ 39

Author(s):

Aaron J. Carman ◽

Slavena Vylkova ◽

Michael C. Lorenz

Keyword(s):

Fatty Acids ◽

Candida Albicans ◽

Coenzyme A ◽

Glyoxylate Cycle ◽

Functional Divergence ◽

Carbon Sources ◽

Growth Defect ◽

Acetyl Coa ◽

Acetyl Coenzyme A ◽

Acetyl Coenzyme

ABSTRACT Acetyl coenzyme A (acetyl-CoA) is the central intermediate of the pathways required to metabolize nonfermentable carbon sources. Three such pathways, i.e., gluconeogenesis, the glyoxylate cycle, and β-oxidation, are required for full virulence in the fungal pathogen Candida albicans. These processes are compartmentalized in the cytosol, mitochondria, and peroxosomes, necessitating transport of intermediates across intracellular membranes. Acetyl-CoA is trafficked in the form of acetate by the carnitine shuttle, and we hypothesized that the enzymes that convert acetyl-CoA to/from acetate, i.e., acetyl-CoA hydrolase (ACH1) and acetyl-CoA synthetase (ACS1 and ACS2), would regulate alternative carbon utilization and virulence. We show that C. albicans strains depleted for ACS2 are unviable in the presence of most carbon sources, including glucose, acetate, and ethanol; these strains metabolize only fatty acids and glycerol, a substantially more severe phenotype than that of Saccharomyces cerevisiae acs2 mutants. In contrast, deletion of ACS1 confers no phenotype, though it is highly induced in the presence of fatty acids, perhaps explaining why acs2 mutants can utilize fatty acids. Strains lacking ACH1 have a mild growth defect on some carbon sources but are fully virulent in a mouse model of disseminated candidiasis. Both ACH1 and ACS2 complement mutations in their S. cerevisiae homolog. Together, these results show that acetyl-CoA metabolism and transport are critical for growth of C. albicans on a wide variety of nutrients. Furthermore, the phenotypic differences between mutations in these highly conserved genes in S. cerevisiae and C. albicans support recent findings that significant functional divergence exists even in fundamental metabolic pathways between these related yeasts.

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Mitochondrial Carnitine-Dependent Acetyl Coenzyme A Transport Is Required for Normal Sexual and Asexual Development of the Ascomycete Gibberella zeae

Eukaryotic Cell ◽

10.1128/ec.00104-12 ◽

2012 ◽

Vol 11 (9) ◽

pp. 1143-1153 ◽

Cited By ~ 17

Author(s):

Hokyoung Son ◽

Kyunghun Min ◽

Jungkwan Lee ◽

Gyung Ja Choi ◽

Jin-Cheol Kim ◽

...

Keyword(s):

Coenzyme A ◽

Developmental Stages ◽

Pathogenic Fungus ◽

Gibberella Zeae ◽

Asexual Development ◽

Acetyl Coa ◽

Alternative Source ◽

Acetyl Coenzyme A ◽

Content Type ◽

Acetyl Coenzyme

ABSTRACTFungi have evolved efficient metabolic mechanisms for the exact temporal (developmental stages) and spatial (organelles) production of acetyl coenzyme A (acetyl-CoA). We previously demonstrated mechanistic roles of several acetyl-CoA synthetic enzymes, namely, ATP citrate lyase and acetyl-CoA synthetases (ACSs), in the plant-pathogenic fungusGibberella zeae. In this study, we characterized two carnitine acetyltransferases (CATs; CAT1 and CAT2) to obtain a better understanding of the metabolic processes occurring inG. zeae. We found that CAT1 functioned as an alternative source of acetyl-CoA required for lipid accumulation in anACS1deletion mutant. Moreover, deletion ofCAT1and/orCAT2resulted in various defects, including changes to vegetative growth, asexual/sexual development, trichothecene production, and virulence. Although CAT1 is associated primarily with peroxisomal CAT function, mislocalization experiments showed that the role of CAT1 in acetyl-CoA transport between the mitochondria and cytosol is important for sexual and asexual development inG. zeae. Taking these data together, we concluded thatG. zeaeCATs are responsible for facilitating the exchange of acetyl-CoA across intracellular membranes, particularly between the mitochondria and the cytosol, during various developmental stages.

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