title: Reduced methylation of PFKFB3 in cancer cells shunts glucose towards the pentose phosphate pathway.
abstract: Haem oxygenase (HO)-1/carbon monoxide (CO) protects cancer cells from oxidative stress, but the gas-responsive signalling mechanisms remain unknown. Here we show using metabolomics that CO-sensitive methylation of PFKFB3, an enzyme producing fructose 2,6-bisphosphate (F-2,6-BP), serves as a switch to activate phosphofructokinase-1, a rate-limiting glycolytic enzyme. In human leukaemia U937 cells, PFKFB3 is asymmetrically di-methylated at R131 and R134 through modification by protein arginine methyltransferase 1. HO-1 induction or CO results in reduced methylation of PFKFB3 in varied cancer cells to suppress F-2,6-BP, shifting glucose utilization from glycolysis toward the pentose phosphate pathway. Loss of PFKFB3 methylation depends on the inhibitory effects of CO on haem-containing cystathionine β-synthase (CBS). CBS modulates remethylation metabolism, and increases NADPH to supply reduced glutathione, protecting cells from oxidative stress and anti-cancer reagents. Once the methylation of PFKFB3 is reduced, the protein undergoes polyubiquitination and is degraded in the proteasome. These results suggest that the CO/CBS-dependent regulation of PFKFB3 methylation determines directional glucose utilization to ensure resistance against oxidative stress for cancer cell survival.
authors.affiliation: Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Naoharu
authors.initials: N
authors.lastname: Takano
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Ishiwata
authors.affiliation: Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Mitsuyo
authors.initials: M
authors.lastname: Ohmura
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Nagahata
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Matsuura
authors.affiliation: Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Aki
authors.initials: A
authors.lastname: Kamata
authors.affiliation: Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Sakamoto
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] MS Business Unit, Shimadzu Corporation, Kyoto 604-8511, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Nakanishi
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Kubo
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Hishiki
authors.affiliation: 1] Department of Biochemistry, Keio University School of Medicine, Tokyo 160-8582, Japan [2] Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Tokyo 160-8582, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Suematsu
mesh_terms: D045744:Cell Line, Tumor; D015536:Down-Regulation; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008745:Methylation; D009369:Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D010427:Pentose Phosphate Pathway; D025481:Phosphofructokinase-2 / Q000235:genetics / Q000378:metabolism*
chemical_list: D006419:Heme Oxygenase (Decyclizing); C107345:PFKFB3 protein, human; D025481:Phosphofructokinase-2; D005947:Glucose