Both antisense and sense expression of biotin carboxyl carrier protein isoform 2 inactivates the plastid acetyl‐coenzyme A carboxylase in <i>Arabidopsis thaliana</i>

Thelen, Jay J.; Ohlrogge, John B.

doi:10.1046/j.1365-313x.2002.01435.x

Cited by 87 publications

(81 citation statements)

References 31 publications

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“…Natural examples of pseudo-poison complex regulation are infrequent, though at least one bioengineered example has been reported. Overexpressing BCCP2 in Arabidopsis seed markedly reduced both hetACCase activity and seed oil content due to increased levels of apo-BCCP (Thelen and Ohlrogge, 2002c;Chen et al, 2009). Pseudo-poisoned hetACCase complexes were formed, reducing carbon flux through de novo FAS.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, reduction of BADC expression in planta might increase hetACCase activity. Since ACCase activity has previously been shown to correlate with seed oil content in transgenic lines altered in ACCase function (Roesler et al, 1997;Thelen and Ohlrogge, 2002c), RNAi silencing of BADC1 in Arabidopsis using a seed-specific promoter was performed. Seed oil content analysis showed a significant increase in three of six independent T2 seeds, as normalized to either seed mass or individual seed ( Figures 9A and 9B).…”

Section: Seed-specific Rnai Silencing Of Badc1 Increases Seed Oil Conmentioning

confidence: 99%

“…Antibodies used in this study were derived from rabbits immunized with recombinant Pisum sativum a-CT (Thelen and Ohlrogge, 2002b), recombinant Arabidopsis BCCP2 (Thelen and Ohlrogge, 2002c), recombinant Arabidopsis BADC2 short peptide (Chen et al, 2009), or recombinant Arabidopsis BADC1 (this study). Blots were probed in secondary antibody for 1 h at room temperature and developed as described previously (Thelen and Ohlrogge, 2002b).…”

Section: Immunoblottingmentioning

confidence: 99%

“…HetACCase activity was directly monitored in 10-d-old siliques or 21-d-old leaves through the incorporation of 14 CO 2 into acid-stable products as described (Thelen and Ohlrogge, 2002c). Details can be found in the Supplemental Methods.…”

Section: Accase Activity Assaysmentioning

confidence: 99%

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A Family of Negative Regulators Targets the Committed Step of de Novo Fatty Acid Biosynthesis

et al. 2016

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ORCID ID: 0000-0002-9686-9645 (V.L.)Acetyl-CoA carboxylase (ACCase) catalyzes the committed step of de novo fatty acid biosynthesis. In prokaryotes, green algae, and most plants, this enzyme is a heteromeric complex requiring four different subunits for activity. The plant complex is recalcitrant to conventional purification schemes and hence the structure and composition of the full assembly have been unclear. In vivo coimmunoprecipitation using subunit-specific antibodies identified a novel family of proteins in Arabidopsis thaliana annotated as biotin/lipoyl attachment domain containing (BADC) proteins. Results from yeast two-hybrid and coexpression in Escherichia coli confirmed that all three BADC isoforms interact with the two biotin carboxyl carrier protein (BCCP) isoforms of Arabidopsis ACCase. These proteins resemble BCCP subunits but are not biotinylated due to a mutated biotinylation motif. We demonstrate that BADC proteins significantly inhibit ACCase activity in both E. coli and Arabidopsis. Targeted gene silencing of BADC isoform 1 in Arabidopsis significantly increased seed oil content when normalized to either mass or individual seed. We conclude the BADC proteins are ancestral BCCPs that gained a new function as negative regulators of ACCase after initial loss of the biotinylation motif. A functional model is proposed.

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Section: Discussionmentioning

confidence: 99%

Section: Seed-specific Rnai Silencing Of Badc1 Increases Seed Oil Conmentioning

confidence: 99%

Section: Immunoblottingmentioning

confidence: 99%

Section: Accase Activity Assaysmentioning

confidence: 99%

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A Family of Negative Regulators Targets the Committed Step of de Novo Fatty Acid Biosynthesis

et al. 2016

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“…The role of biotin carboxyl carrier protein in fatty acids deposition in seeds was characterized by transgenic Arabidopsis thaliana plants of up-and down-activated gene for BCCP2 -an isoform predominantly expressed in flowers and developing seeds (Thelen and Ohlrogge, 2002). Down-regulation of this gene did not reveal a significant phenotype apart from a reduced level of the protein and a decreased, by 9%, content of fatty acids in seeds.…”

Section: Accase -Essential Enzyme For Oil Production In Plantsmentioning

confidence: 99%

OPINIONS Acetyl-coenzyme A carboxylase – an attractive enzyme for biotechnology

Podkowiński¹,

Tworak²

2011

bta

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Acetyl-CoA carboxylase catalyzes the carboxylation of acetyl-CoA to malonyl-CoA. This reaction constitutes the first committed step of fatty acid synthesis in most organisms, while its product also serves as a universal precursor for various other high-value compounds. The important regulatory and rate-limiting role of acetyl-CoA carboxylase makes this enzyme a powerful tool in a variety of biotechnological and medical projects. This review presents the current knowledge on structural and functional features of the enzyme and focuses on its divergent applications. Different metabolic engineering attempts that lead to the production of various compounds, such as fatty acids, polyketides or flavonoids, are presented and their advantages and limitations discussed. The importance of studies on acetyl-CoA carboxylase activity for design and development of new herbicides and antibiotics as well as for medical intervention is also highlighted. Acetyl-coenzyme A carboxylase -enzyme architecture, biochemistry and its role in cell physiologyA variety of biotechnological projects targeted to modulate acetyl-coenzyme A carboxylase activity in bacteria, plants and humans have been proposed and experimentally tested. Here, we would like to present a comprehensive review of these strategies together with a survey of the potential of acetyl-CoA carboxylase for biotechnology.Acetyl-coenzyme A carboxylase (ACC, E.C.6.4.1.2), recognized as an essential enzyme for all Eukaryotes and the majority of Bacteria, is responsible for carboxylation of acetyl-CoA that results in malonyl-coenzyme A formation (Fig. 1). Malonyl-CoA is a major building block for compounds such as fatty acids, polyketides and flavonoids -important components for cell growth, as well as for food, pharmaceuticals and energy production.The malonyl-CoA synthesis consists of two half-reactions (Nikolau et al., 2003). The first one, adenosine triphosphate-dependent carboxylation of biotin prosthetic group covalently bound to a module named biotin carboxyl carrier protein (BCCP), is catalyzed by ACCase segment of biotin carboxylase activity (BC) (Fig. 2). This reaction is immediately followed by the second half-re action: the transfer of a carboxyl group from carboxylated biotin to acetyl-CoA, resulting in malonyl-CoA formation. Carboxyl transferase (CT) is a module that catalyzes the second reaction and provides the required specificity in recognition of the carboxyl acceptor (acetyl-CoA). The ACCase model usually presents BCCP, a module with covalently attached biotin, as a beam responsible for biotin dislocation between two active centers -one with BT and the other with CT activity. The three (Fig. 3). The prokaryotic and eukaryotic types of ACCases are evolutionary related and the phylogeny of the modules provides some hints about their cenancestor, a split between Archaea and Bacteria, and arising of Eukaryota (Lombard and Moreira, 2011).Phylogeny is out of scope of this manuscript, but acetyl-coenzyme A carboxylases from various organisms representing v...

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Comparative proteomics of seed maturation in oilseeds reveals differences in intermediary metabolism

et al. 2011

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Proteomics is increasingly being used to understand enzyme expression and regulatory mechanisms involved in the accumulation of storage reserves in crops with sequenced genomes. During the past six years, considerable progress has been made to characterize proteomes of both mature and developing seeds, particularly oilseeds - plants which accumulate principally oil and protein as storage reserves. This review summarizes the emerging proteomics data, with emphasis on seed filling in soy, rapeseed, castor and Arabidopsis as each of these oilseeds were analyzed using very similar proteomic strategies. These parallel studies provide a comprehensive view of source-sink relationships, specifically sucrose assimilation into organic acid intermediates for de novo amino acid and fatty acid synthesis. We map these biochemical processes for seed maturation and illustrate the differences and similarities among the four oilseeds. For example, while the four oilseeds appear capable of producing cytosolic phosphoenolpyruvate as the principal carbon intermediate, soybean and castor also express malic enzymes and malate dehydrogenase, together capable of producing malate that has been previously proposed to be the major intermediate for fatty acid synthesis in castor. We discuss these and other differences in the context of intermediary metabolism for developing oilseeds.

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Both antisense and sense expression of biotin carboxyl carrier protein isoform 2 inactivates the plastid acetyl‐coenzyme A carboxylase in Arabidopsis thaliana

Cited by 87 publications

References 31 publications

A Family of Negative Regulators Targets the Committed Step of de Novo Fatty Acid Biosynthesis

A Family of Negative Regulators Targets the Committed Step of de Novo Fatty Acid Biosynthesis

OPINIONS Acetyl-coenzyme A carboxylase – an attractive enzyme for biotechnology

Comparative proteomics of seed maturation in oilseeds reveals differences in intermediary metabolism

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