Quantitative Analysis of Purine Nucleotides Indicates That Purinosomes Increase de Novo Purine Biosynthesis

Zhao, Hong; Chiaro, Christopher; Zhang, Limin; Smith, Philip B.; Chan, Collin; Pedley, Anthony M.; Pugh, Raymond; French, Jarrod B.; Patterson, Andrew D.; Benkovic, Stephen J.

doi:10.1074/jbc.m114.628701

Cited by 109 publications

(140 citation statements)

References 37 publications

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“…The concept of the metabolon has stirred up more than a little heat, with much expectation but little experimental verification [53]. More recently, experimental evidence for metabolons has appeared [54], including evidence for a glycolytic metabolon in plants [55], and it is becoming clear that metabolons do exist but are dynamic rather than static, being induced when there is a metabolic need, and they almost always assemble on membranes (or on other surfaces such as the cytoskeleton) [56]. In erythrocytes, glycolytic enzymes associate with the anion transporter band 3, in areas where ATP is consumed, forming an ordered complex [57,58].…”

Section: Discussionmentioning

confidence: 99%

Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate

Yasid¹,

Rolfe²,

Green³

et al. 2016

R. Soc. open sci.

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We have developed a method for rapid quenching of samples taken from chemostat cultures of Escherichia coli that gives reproducible and reliable measurements of extracellular and intracellular metabolites by 1H NMR and have applied it to study the major central metabolites during the transition from anaerobic to aerobic growth. Almost all metabolites showed a gradual change after perturbation with air, consistent with immediate inhibition of pyruvate formate-lyase, dilution of overflow metabolites and induction of aerobic enzymes. Surprisingly, although pyruvate showed almost no change in intracellular concentration, the extracellular concentration transiently increased. The absence of intracellular accumulation of pyruvate suggested that one or more glycolytic enzymes might relocate to the cell membrane. To test this hypothesis, chromosomal pyruvate kinase (pykF) was modified to express either PykF-green fluorescent protein or PykF-FLAG fusion proteins. Measurements showed that PykF-FLAG relocates to the cell membrane within 5 min of aeration and then slowly returns to the cytoplasm, suggesting that on aeration, PykF associates with the membrane to facilitate secretion of pyruvate to maintain constant intracellular levels.

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Section: Discussionmentioning

confidence: 99%

Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate

Yasid¹,

Rolfe²,

Green³

et al. 2016

R. Soc. open sci.

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“…Probing with the remaining four pathway enzymes also showed co-clustering with FGAMS [6]. Immunofluorescence has demonstrated purinosome formation on the endogenous level and provided evidence that the observed compartmentalization is not a consequence of overexpression due to transient transfection [6, 34-36]. Additionally, particle characterization in transient transfected models demonstrated that these enzyme clusters are distinct in size and cell density from processing bodies (P-bodies), stress granules, and aggresomes (Box 2) [37].…”

Section: Discovery Of a Metabolon In Purine Metabolism – The Purinosomementioning

confidence: 99%

A New View into the Regulation of Purine Metabolism: The Purinosome

Pedley

Benkovic

2017

Trends in Biochemical Sciences

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429

361

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Other than serving as building blocks for DNA and RNA, purines metabolites provide a cell with the necessary energy and cofactors to promote cell survival and proliferation. A renewed interest in how purine metabolism may fuel cancer progression has uncovered a new perspective into how a cell regulates purine need. Under cellular conditions of high purine demand, the de novo purine biosynthetic enzymes cluster near mitochondria and microtubules to form dynamic multi-enzyme complexes referred to as purinosomes. This review highlights the purinosome as a novel level of metabolic organization of enzymes in cells, its consequences for regulation of purine metabolism, and the extent that purine metabolism is being targeted for the treatment of cancers.

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“…Purinosome formation and dissociation were found to be modulated by several factors, including the microtubule network and cell signaling through protein phosphorylation (Deng et al, 2012). Significantly, although IMPDH itself appears to be a purinosome component, RR and purinosomes were shown to be distinct cellular bodies, which, under favorable conditions, could be simultaneously visualized within the same cells (Zhao et al, 2015). One cannot dismiss the possibility that RRs form when IMPDH levels required by the cell exceed the amount needed to form purinosome complexes.…”

Section: Discussionmentioning

confidence: 99%

“…Manual counting was performed using the Cell Counter plugin included in Fiji or ImageJ software (Schindelin et al, 2012). With the knowledge that IMPDH localizes to purinosomes as well as RRs (Zhao et al, 2015), only mature filamentous RRs were included in the quantification of the percentage of cells containing RRs. Any foci or short, immature RRs that could not be readily distinguished from purinosomes or other assemblies were not considered RRs.…”

Section: Statistical Analysis and Quantification Of Rrsmentioning

confidence: 99%

‘Rod and ring’ formation from IMP dehydrogenase is regulated through the one-carbon metabolic pathway

Calise

Purich

Nguyen

et al. 2016

Journal of Cell Science

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ABSTRACT'Rods and rings' (RRs) are conserved, non-membrane-bound intracellular polymeric structures composed, in part, of inosine monophosphate dehydrogenase (IMPDH), a key enzyme leading to GMP and GTP biosynthesis. RR formation is induced by IMPDH inhibitors as well as glutamine deprivation. They also form upon treatment of cells with glutamine synthetase inhibitors. We now report that depriving cells of serine and glycine promotes RR formation, and we have traced these effects to dihydrofolate reductase (DHFR) and serine hydroxymethyltransferase-2 (SHMT2), pivotal enzymes in one-carbon metabolism and nucleotide biosynthesis. RR assembly is likewise induced upon DHFR inhibition by methotrexate or aminopterin as well as siRNA-mediated knockdown of DHFR or SHMT2. Because RR assembly occurs when guanine nucleotide biosynthesis is inhibited, and because RRs rapidly disassemble after the addition of guanine nucleotide precursors, RR formation might be an adaptive homeostatic mechanism, allowing IMPDH to sense changes in the one-carbon folate pathway.

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Quantitative Analysis of Purine Nucleotides Indicates That Purinosomes Increase de Novo Purine Biosynthesis

Cited by 109 publications

References 37 publications

Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate

Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate

A New View into the Regulation of Purine Metabolism: The Purinosome

‘Rod and ring’ formation from IMP dehydrogenase is regulated through the one-carbon metabolic pathway

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