Mammalian TAK1 activates Snf1 protein kinase in yeast and phosphorylates AMP-activated protein kinase in vitro.
The Snf1/AMP-activated protein kinase (AMPK) family is important for metabolic regulation and is highly conserved from yeast to mammals. The upstream kinases are also functionally conserved, and the AMPK kinases LKB1 and Ca2+/calmodulin-dependent protein kinase kinase activate Snf1 in mutant yeast cells lacking the native Snf1-activating kinases, Sak1, Tos3, and Elm1. Here, we exploited the yeast genetic system to identify members of the mammalian AMPK kinase family by their function as Snf1-activating kinases. A mouse embryo cDNA library in a yeast expression vector was used to transform sak1Delta tos3Delta elm1Delta yeast cells. Selection for a Snf+ growth phenotype yielded cDNA plasmids expressing LKB1, Ca2+/calmodulin-dependent protein kinase kinase, and transforming growth factor-beta-activated kinase (TAK1), a member of the mitogen-activated protein kinase kinase kinase family. We present genetic and biochemical evidence that TAK1 activates Snf1 protein kinase in vivo and in vitro. We further show that recombinant TAK1, fused to the activation domain of its binding partner TAB1, phosphorylates Thr-172 in the activation loop of the AMPK catalytic domain. Finally, expression of TAK1 and TAB1 in HeLa cells or treatment of cells with cytokines stimulated phosphorylation of Thr-172 of AMPK. These findings indicate that TAK1 is a functional member of the Snf1/AMPK kinase family and support TAK1 as a candidate for an authentic AMPK kinase in mammalian cells.
Momcilovic
Milica
M
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA.
Hong
Seung-Pyo
SP
Carlson
Marian
M
D055372:AMP-Activated Protein Kinases; D000818:Animals; D002118:Calcium / Q000378:metabolism; D017124:Conserved Sequence; D004789:Enzyme Activation; D005656:Fungal Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism / Q000502:physiology*; D009097:Multienzyme Complexes / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D013912:Threonine / Q000737:chemistry; D020798:Two-Hybrid System Techniques; D000099151:MAP Kinase Kinase Kinase 7
D055372:AMP-Activated Protein Kinases; D002118:Calcium; D005656:Fungal Proteins; D020930:MAP Kinase Kinase Kinases; D009097:Multienzyme Complexes; D017346:Protein Serine-Threonine Kinases; D011994:Recombinant Proteins; D013912:Threonine; D000099151:MAP Kinase Kinase Kinase 7; C071570:SNF1-related protein kinases