Metabolite and post-translational control of phosphoenolpyruvate carboxylase from leaves and mesophyll cell protoplasts of Arabidopsis thaliana

Gousset-Dupont, Aurélie; Lebouteiller, Bénédicte; Monreal, J. A.; Echevarrı́a, Cristina; Pierre, Jean‐Noël; Hodges, Michael; Vidal, Jean

doi:10.1016/j.plantsci.2005.07.014

Cited by 22 publications

(21 citation statements)

References 24 publications

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“…Thus, our results indicated that PPC1 and PPC2 were the most highly expressed PEPC genes in the leaves. These results confirmed the findings of several earlier Arabidopsis PEPC studies (Gousset-Dupont et al, 2005;Gregory et al, 2009).…”

supporting

confidence: 92%

Phosphoenolpyruvate Carboxylase in Arabidopsis Leaves Plays a Crucial Role in Carbon and Nitrogen Metabolism

et al. 2015

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Phosphoenolpyruvate carboxylase (PEPC) is a crucial enzyme that catalyzes an irreversible primary metabolic reaction in plants. Previous studies have used transgenic plants expressing ectopic PEPC forms with diminished feedback inhibition to examine the role of PEPC in carbon and nitrogen metabolism. To date, the in vivo role of PEPC in carbon and nitrogen metabolism has not been analyzed in plants. In this study, we examined the role of PEPC in plants, demonstrating that PPC1 and PPC2 were highly expressed genes encoding PEPC in Arabidopsis (Arabidopsis thaliana) leaves and that PPC1 and PPC2 accounted for approximately 93% of total PEPC activity in the leaves. A double mutant, ppc1/ppc2, was constructed that exhibited a severe growth-arrest phenotype. The ppc1/ppc2 mutant accumulated more starch and sucrose than wild-type plants when seedlings were grown under normal conditions. Physiological and metabolic analysis revealed that decreased PEPC activity in the ppc1/ppc2 mutant greatly reduced the synthesis of malate and citrate and severely suppressed ammonium assimilation. Furthermore, nitrate levels in the ppc1/ppc2 mutant were significantly lower than those in wild-type plants due to the suppression of ammonium assimilation. Interestingly, starch and sucrose accumulation could be prevented and nitrate levels could be maintained by supplying the ppc1/ppc2 mutant with exogenous malate and glutamate, suggesting that low nitrogen status resulted in the alteration of carbon metabolism and prompted the accumulation of starch and sucrose in the ppc1/ppc2 mutant. Our results demonstrate that PEPC in leaves plays a crucial role in modulating the balance of carbon and nitrogen metabolism in Arabidopsis.

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supporting

confidence: 92%

Phosphoenolpyruvate Carboxylase in Arabidopsis Leaves Plays a Crucial Role in Carbon and Nitrogen Metabolism

et al. 2015

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“…Limited proteolytic degradation of p118 occurred during co-IP at room temperature, despite our use of conditions documented to negate its proteolysis during incubation of clarified COS extracts on ice (i.e. absence of thiol reducing agents, inclusion of ProteCEASE-100; Gennidakis et al, 2007 laser desorption time-of-flight (MALDI-TOF) MS analysis of tryptic peptides derived from the solubilized immunoprecipitate indicated the presence of all three Arabidopsis PTPC isozymes (AtPPC1-AtPPC3), in addition to the AtPPC4 BTPC (Gousset-Dupont et al, 2005).…”

Section: Btpc Quantitatively Coimmuopurifies With Ptpc From Developinmentioning

confidence: 99%

Coimmunopurification of Phosphorylated Bacterial- and Plant-Type Phosphoenolpyruvate Carboxylases with the Plastidial Pyruvate Dehydrogenase Complex from Developing Castor Oil Seeds

et al. 2008

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The phosphoenolpyruvate carboxylase (PEPC) interactome of developing castor oil seed (COS; Ricinus communis) endosperm was assessed using coimmunopurification (co-IP) followed by proteomic analysis. Earlier studies suggested that immunologically unrelated 107-kD plant-type PEPCs (p107/PTPC) and 118-kD bacterial-type PEPCs (p118/BTPC) are subunits of an unusual 910-kD hetero-octameric class 2 PEPC complex of developing COS. The current results confirm that a tight physical interaction occurs between p118 and p107 because p118 quantitatively coimmunopurified with p107 following elution of COS extracts through an anti-p107-IgG immunoaffinity column. No PEPC activity or immunoreactive PEPC polypeptides were detected in the corresponding flow-through fractions. Although BTPCs lack the N-terminal phosphorylation motif characteristic of PTPCs, Pro-Q Diamond phosphoprotein staining, immunoblotting with phospho-serine (Ser)/threonine Akt substrate IgG, and phosphate-affinity PAGE established that coimmunopurified p118 was multiphosphorylated at unique Ser and/or threonine residues. Tandem mass spectrometric analysis of an endoproteinase Lys-C p118 peptide digest demonstrated that Ser-425 is subject to in vivo proline-directed phosphorylation. The co-IP of p118 with p107 did not appear to be influenced by their phosphorylation status. Because p118 phosphorylation was unchanged 48 h following elimination of photosynthate supply due to COS depodding, the signaling mechanisms responsible for photosynthate-dependent p107 phosphorylation differ from those controlling p118's in vivo phosphorylation. A 110-kD PTPC coimmunopurified with p118 and p107 when depodded COS was used. The plastidial pyruvate dehydrogenase complex (PDC pl ) was identified as a novel PEPC interactor. Thus, a putative metabolon involving PEPC and PDC pl could function to channel carbon from phosphoenolpyruvate to acetyl-coenzyme A and/or to recycle CO 2 from PDC pl to PEPC.

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“…PEPC kinase enzyme is mainly regulated by transcriptional level abundance, showing the peculiar characteristic among kinase enzymes of requiring de novo synthesis to be active [55]. In C 3 and C 4 plants, PPCK seems to be activated by light [56][57][58][59], while in CAM plants this enzyme responds mostly to a circadian oscillator and is active during the night [55,60,62]. Taybi and cols.…”

Section: An Overview Of Pepc and Ppck Regulationmentioning

confidence: 99%

CAM Photosynthesis in Bromeliads and Agaves: What Can We Learn from These Plants?

Matiz¹,

Mioto²,

Mayorga³

et al. 2013

Photosynthesis

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Metabolite and post-translational control of phosphoenolpyruvate carboxylase from leaves and mesophyll cell protoplasts of Arabidopsis thaliana

Cited by 22 publications

References 24 publications

Phosphoenolpyruvate Carboxylase in Arabidopsis Leaves Plays a Crucial Role in Carbon and Nitrogen Metabolism

Phosphoenolpyruvate Carboxylase in Arabidopsis Leaves Plays a Crucial Role in Carbon and Nitrogen Metabolism

Coimmunopurification of Phosphorylated Bacterial- and Plant-Type Phosphoenolpyruvate Carboxylases with the Plastidial Pyruvate Dehydrogenase Complex from Developing Castor Oil Seeds

CAM Photosynthesis in Bromeliads and Agaves: What Can We Learn from These Plants?

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