title: Tyr-301 phosphorylation inhibits pyruvate dehydrogenase by blocking substrate binding and promotes the Warburg effect.
abstract: The mitochondrial pyruvate dehydrogenase complex (PDC) plays a crucial role in regulation of glucose homoeostasis in mammalian cells. PDC flux depends on catalytic activity of the most important enzyme component pyruvate dehydrogenase (PDH). PDH kinase inactivates PDC by phosphorylating PDH at specific serine residues, including Ser-293, whereas dephosphorylation of PDH by PDH phosphatase restores PDC activity. The current understanding suggests that Ser-293 phosphorylation of PDH impedes active site accessibility to its substrate pyruvate. Here, we report that phosphorylation of a tyrosine residue Tyr-301 also inhibits PDH α 1 (PDHA1) by blocking pyruvate binding through a novel mechanism in addition to Ser-293 phosphorylation. In addition, we found that multiple oncogenic tyrosine kinases directly phosphorylate PDHA1 at Tyr-301, and Tyr-301 phosphorylation of PDHA1 is common in EGF-stimulated cells as well as diverse human cancer cells and primary leukemia cells from human patients. Moreover, expression of a phosphorylation-deficient PDHA1 Y301F mutant in cancer cells resulted in increased oxidative phosphorylation, decreased cell proliferation under hypoxia, and reduced tumor growth in mice. Together, our findings suggest that phosphorylation at distinct serine and tyrosine residues inhibits PDHA1 through distinct mechanisms to impact active site accessibility, which act in concert to regulate PDC activity and promote the Warburg effect.
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322,. Electronic address: jfan3@emory.edu.
authors.forename: Jun
authors.initials: J
authors.lastname: Fan
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Hee-Bum
authors.initials: HB
authors.lastname: Kang
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Changliang
authors.initials: C
authors.lastname: Shan
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Shannon
authors.initials: S
authors.lastname: Elf
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Ruiting
authors.initials: R
authors.lastname: Lin
authors.affiliation: Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, and.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Xie
authors.affiliation: Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, and.
authors.forename: Ting-Lei
authors.initials: TL
authors.lastname: Gu
authors.affiliation: Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, and.
authors.forename: Mike
authors.initials: M
authors.lastname: Aguiar
authors.affiliation: Cell Signaling Technology, Inc., Danvers, Massachusetts 01923, and.
authors.forename: Scott
authors.initials: S
authors.lastname: Lonning
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Tae-Wook
authors.initials: TW
authors.lastname: Chung
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Martha
authors.initials: M
authors.lastname: Arellano
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Hanna J
authors.initials: HJ
authors.lastname: Khoury
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Dong M
authors.initials: DM
authors.lastname: Shin
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Fadlo R
authors.initials: FR
authors.lastname: Khuri
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520.
authors.forename: Titus J
authors.initials: TJ
authors.lastname: Boggon
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: Sumin
authors.initials: S
authors.lastname: Kang
authors.affiliation: Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory, Emory University School of Medicine, Atlanta, Georgia 30322,. Electronic address: jchen@emory.edu.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
mesh_terms: D016475:3T3 Cells; D019943:Amino Acid Substitution; D000818:Animals; D050260:Carbohydrate Metabolism; D020134:Catalytic Domain; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004815:Epidermal Growth Factor / Q000502:physiology; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010085:Oxidative Phosphorylation; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D030481:Pyruvate Dehydrogenase (Lipoamide) / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019289:Pyruvic Acid / Q000737:chemistry; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000378:metabolism; D047368:Tumor Burden; D014443:Tyrosine / Q000378:metabolism
chemical_list: D014443:Tyrosine; D004815:Epidermal Growth Factor; D019289:Pyruvic Acid; D030481:Pyruvate Dehydrogenase (Lipoamide); C065640:pyruvate dehydrogenase E1alpha subunit; C496348:FGFR1 protein, human; D051496:Receptor, Fibroblast Growth Factor, Type 1
keywords: Cell Proliferation; Phosphotyrosine Signaling; Pyruvate Dehydrogenase Complex (PDC); Tumor Metabolism; Warburg Effect
references.citation: Adv Enzyme Regul. 2002;42:249-59
references.pmid: 12123719
references.citation: Blood. 2004 Apr 1;103(7):2802-5
references.pmid: 14630792
references.citation: Cell. 2006 Aug 25;126(4):663-76
references.pmid: 16904174
references.citation: Cancer Res. 2004 Jun 1;64(11):3892-9
references.pmid: 15172999
references.citation: J Cell Biochem Suppl. 1996;24:1-291
references.pmid: 8965489
references.citation: Blood. 2006 Dec 15;108(13):4202-4
references.pmid: 16946300
references.citation: Mol Cell. 2014 Feb 20;53(4):534-48
references.pmid: 24486017
references.citation: Leukemia. 1997 Sep;11(9):1469-77
references.pmid: 9305600
references.citation: Sci Signal. 2009 Nov 17;2(97):ra73
references.pmid: 19920251
references.citation: Amino Acids. 2009 Mar;36(3):449-56
references.pmid: 18500430
references.citation: Blood. 1983 Oct;62(4):832-45
references.pmid: 6309287
references.citation: Cancer Res. 2006 Sep 15;66(18):8927-30
references.pmid: 16982728
references.citation: Clin Cancer Res. 2004 Oct 1;10(19):6661-8
references.pmid: 15475456
references.citation: J Natl Cancer Inst. 1973 Nov;51(5):1417-23
references.pmid: 4357758
references.citation: Nat Commun. 2013;4:1790
references.pmid: 23653202
references.citation: J Natl Cancer Inst. 1973 Nov;51(5):1409-16
references.pmid: 4357757
references.citation: J Natl Cancer Inst. 1974 Sep;53(3):661-74
references.pmid: 4412247
references.citation: J Invest Dermatol. 1999 Mar;112(3):343-53
references.pmid: 10084313
references.citation: Mol Cell Biol. 2011 Dec;31(24):4938-50
references.pmid: 21969607
references.citation: Cell. 2002 Jul 12;110(1):19-32
references.pmid: 12150994
references.citation: Nat Rev Cancer. 2011 Feb;11(2):85-95
references.pmid: 21258394
references.citation: Acta Crystallogr D Biol Crystallogr. 2001 Oct;57(Pt 10):1373-82
references.pmid: 11567148
references.citation: Blood. 1975 Mar;45(3):321-34
references.pmid: 163658
references.citation: Cancer Cell. 2008 Jun;13(6):472-82
references.pmid: 18538731
references.citation: Cell Metab. 2006 Mar;3(3):177-85
references.pmid: 16517405
references.citation: J Biol Chem. 2003 Jun 6;278(23):21240-6
references.pmid: 12651851
references.citation: Prog Nucleic Acid Res Mol Biol. 2001;70:33-75
references.pmid: 11642366
references.citation: J Biol Chem. 1994 Sep 23;269(38):23757-63
references.pmid: 8089148
references.citation: J Biol Chem. 2014 Aug 1;289(31):21413-22
references.pmid: 24962578
references.citation: Mol Cell. 2011 Dec 23;44(6):864-77
references.pmid: 22195962
references.citation: Biochemistry. 2007 May 29;46(21):6277-87
references.pmid: 17474719
references.citation: J Biol Chem. 2004 Feb 20;279(8):6921-33
references.pmid: 14638692