title: Chemical genetic screen for AMPKα2 substrates uncovers a network of proteins involved in mitosis.
abstract: The energy-sensing AMP-activated protein kinase (AMPK) is activated by low nutrient levels. Functions of AMPK, other than its role in cellular metabolism, are just beginning to emerge. Here we use a chemical genetics screen to identify direct substrates of AMPK in human cells. We find that AMPK phosphorylates 28 previously unidentified substrates, several of which are involved in mitosis and cytokinesis. We identify the residues phosphorylated by AMPK in vivo in several substrates, including protein phosphatase 1 regulatory subunit 12C (PPP1R12C) and p21-activated protein kinase (PAK2). AMPK-induced phosphorylation is necessary for PPP1R12C interaction with 14-3-3 and phosphorylation of myosin regulatory light chain. Both AMPK activity and PPP1R12C phosphorylation are increased in mitotic cells and are important for mitosis completion. These findings suggest that AMPK coordinates nutrient status with mitosis completion, which may be critical for the organism's response to low nutrients during development, or in adult stem and cancer cells.
authors.affiliation: Department of Genetics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Max R
authors.initials: MR
authors.lastname: Banko
authors.forename: Jasmina J
authors.initials: JJ
authors.lastname: Allen
authors.forename: Bethany E
authors.initials: BE
authors.lastname: Schaffer
authors.forename: Erik W
authors.initials: EW
authors.lastname: Wilker
authors.forename: Peiling
authors.initials: P
authors.lastname: Tsou
authors.forename: Jamie L
authors.initials: JL
authors.lastname: White
authors.forename: Judit
authors.initials: J
authors.lastname: Villén
authors.forename: Beatrice
authors.initials: B
authors.lastname: Wang
authors.forename: Sara R
authors.initials: SR
authors.lastname: Kim
authors.forename: Kei
authors.initials: K
authors.lastname: Sakamoto
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: Lewis C
authors.initials: LC
authors.lastname: Cantley
authors.forename: Michael B
authors.initials: MB
authors.lastname: Yaffe
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.forename: Anne
authors.initials: A
authors.lastname: Brunet
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D045744:Cell Line, Tumor; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008938:Mitosis / Q000235:genetics*; D018994:Myosin Light Chains / Q000378:metabolism; D010766:Phosphorylation; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D054462:p21-Activated Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D018994:Myosin Light Chains; D000255:Adenosine Triphosphate; C515907:PAK2 protein, human; C526181:PRKAA2 protein, human; D054462:p21-Activated Kinases; D055372:AMP-Activated Protein Kinases; C540874:PPP1R12C protein, human; D054645:Protein Phosphatase 1