Clem, Brian F.Simmons, Alan J.Lane, Andrew N.Nelson, KristinClem, Amy L.Brock, ErinWattenberg, Brinks W.Telang, SuchetaChesney, Jason2021-10-222021-10-222009-09-04Yalçın, A. vd. (2009). "Nuclear targeting of 6-Phosphofructo-2-kinase (PFKFB3) increases proliferation via cyclin-dependent kinases". Journal of Biological Chemistry, 284(36), 24223-24232.1083-351Xhttps://doi.org/10.1074/jbc.M109.016816https://www.sciencedirect.com/science/article/pii/S0021925819547913http://hdl.handle.net/11452/22431The regulation of metabolism and growth must be tightly coupled to guarantee the efficient use of energy and anabolic substrates throughout the cell cycle. Fructose 2,6-bisphosphate (Fru-2,6-BP) is an allosteric activator of 6-phosphofructo-1-kinase (PFK-1), a rate-limiting enzyme and essential control point in glycolysis. The concentration of Fru-2,6-BP in mammalian cells is set by four 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4), which interconvert fructose 6-phosphate and Fru-2,6-BP. The relative functions of the PFKFB3 and PFKFB4 enzymes are of particular interest because they are activated in human cancers and increased by mitogens and low oxygen. We examined the cellular localization of PFKFB3 and PFKFB4 and unexpectedly found that whereas PFKFB4 localized to the cytoplasm (i.e. the site of glycolysis), PFKFB3 localized to the nucleus. We then overexpressed PFKFB3 and observed no change in glucose metabolism but rather a marked increase in cell proliferation. These effects on proliferation were completely abrogated by mutating either the active site or nuclear localization residues of PFKFB3, demonstrating a requirement for nuclear delivery of Fru-2,6-BP. Using protein array analyses, we then found that ectopic expression of PFKFB3 increased the expression of several key cell cycle proteins, including cyclin-dependent kinase (Cdk)-1, Cdc25C, and cyclinD3 and decreased the expression of the cell cycle inhibitor p27, a universal inhibitor of Cdk-1 and the cell cycle. We also observed that the addition of Fru-2,6-BP to HeLa cell lysates increased the phosphorylation of the Cdk-specific Thr-187 site of p27. Taken together, these observations demonstrate an unexpected role for PFKFB3 in nuclear signaling and indicate that Fru-2,6-BP may couple the activation of glucose metabolism with cell proliferation.eninfo:eu-repo/semantics/openAccessCell-cycleFructose 2,6-bisphosphateLiver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatasecArbohydrate-metabolismBisphosphatase reactionInhibitor p27P27(KIP1)CancerPhosphorylationDegradationBiochemistry & molecular biologyMammaliaBiochemistryCell proliferationChemical activationEnzymesFructoseGlucoseMammalsMetabolismOxygenPathologyActive siteAllosteric activatorBisphosphatesCell cycleCellular localizationConcentration ofControl pointCyclin D3Cyclin-dependent kinaseEctopic expressionsEfficient use of energyGlucose metabolismHeLa cellHuman cancerIn-cellLow oxygenMammalian cellsMitogensNuclear deliveryNuclear localizationNuclear targetingProtein arraysRate-limiting enzymesTightly-coupledCell membranesAllosteric regulationCDC2 protein kinaseCell nucleusCDC25 phosphatasesCell proliferationCyclinsCytoplasmFructosediphosphatesFructosephosphatesGlucoseGlycolysisHela cellsHumansIntracellular signaling peptides and proteinsNeoplasm proteinsNeoplasmsNuclear localization signalsPhosphofructokinase-2Article0002693802000362-s2.0-6994912282824223242322843619473963Biochemistry & molecular biologyPhosphofructokinase-2; Fructose 2,6-Diphosphate; Glycolysis6 phosphofructo 2 kinase fructose 2,6 bisphosphatase 36 phosphofructo 2 kinase fructose 2,6 bisphosphatase 4Cyclin D3Cyclin dependent kinaseCyclin dependent kinase 1Fructose 2,6 bisphosphateGlucosePhosphotransferaseProtein p27Protein tyrosine phosphataseThreonineUnclassified drugArticleCell nucleusCell proliferationCellular distributionControlled studyCytoplasmGene mutationGlucose metabolismHeLa cellHumanHuman cellNucleotide sequencePriority journalProtein expressionProtein functionProtein localizationProtein phosphorylationProtein targetingSignal transduction