Kinome Screen Identifies PFKFB3 and Glucose Metabolism as Important Regulators of the Insulin/Insulin-like Growth Factor (IGF)-1 Signaling Pathway.
The insulin/insulin-like growth factor (IGF)-1 signaling pathway (ISP) plays a fundamental role in long term health in a range of organisms. Protein kinases including Akt and ERK are intimately involved in the ISP. To identify other kinases that may participate in this pathway or intersect with it in a regulatory manner, we performed a whole kinome (779 kinases) siRNA screen for positive or negative regulators of the ISP, using GLUT4 translocation to the cell surface as an output for pathway activity. We identified PFKFB3, a positive regulator of glycolysis that is highly expressed in cancer cells and adipocytes, as a positive ISP regulator. Pharmacological inhibition of PFKFB3 suppressed insulin-stimulated glucose uptake, GLUT4 translocation, and Akt signaling in 3T3-L1 adipocytes. In contrast, overexpression of PFKFB3 in HEK293 cells potentiated insulin-dependent phosphorylation of Akt and Akt substrates. Furthermore, pharmacological modulation of glycolysis in 3T3-L1 adipocytes affected Akt phosphorylation. These data add to an emerging body of evidence that metabolism plays a central role in regulating numerous biological processes including the ISP. Our findings have important implications for diseases such as type 2 diabetes and cancer that are characterized by marked disruption of both metabolism and growth factor signaling.
From the Garvan Institute of Medical Research, Sydney 2010 NSW, Australia.
Sophie
S
Trefely
From the Garvan Institute of Medical Research, Sydney 2010 NSW, Australia, Genentech Inc., South San Francisco, California 94080.
Poh-Sim
PS
Khoo
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and.
James R
JR
Krycer
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and.
Rima
R
Chaudhuri
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and.
Daniel J
DJ
Fazakerley
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and.
Benjamin L
BL
Parker
From the Garvan Institute of Medical Research, Sydney 2010 NSW, Australia.
Ghazal
G
Sultani
Genentech Inc., South San Francisco, California 94080.
James
J
Lee
Genentech Inc., South San Francisco, California 94080.
Jean-Philippe
JP
Stephan
Genentech Inc., South San Francisco, California 94080.
Eric
E
Torres
Genentech Inc., South San Francisco, California 94080.
Kenneth
K
Jung
Genentech Inc., South San Francisco, California 94080.
Coenraad
C
Kuijl
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and the Sydney Medical School, University of Sydney, Sydney 2006 NSW, Australia david.james@sydney.edu.au.
David E
DE
James
Genentech Inc., South San Francisco, California 94080, jagathjr@gmail.com.
Jagath R
JR
Junutula
the Charles Perkins Centre, School of Molecular Bioscience, University of Sydney, Sydney 2006 NSW, Australia, and jacqueline.stoeckli@sydney.edu.au.
Jacqueline
J
Stöckli
D041721:3T3-L1 Cells; D000818:Animals; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4 / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D051379:Mice; D025481:Phosphofructokinase-2 / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction
D051275:Glucose Transporter Type 4; D007328:Insulin; D034741:RNA, Small Interfering; C495692:SLC2A4 protein, human; D007334:Insulin-Like Growth Factor I; D011494:Protein Kinases; C000595376:PFKFB3 protein, mouse; D025481:Phosphofructokinase-2; D005947:Glucose
Akt PKB; PFKFB3; Warburg effect; glucose metabolism; glucose transporter type 4 (GLUT4); glycolysis; insulin