title: Metabolite Regulation of Nuclear Localization of Carbohydrate-response Element-binding Protein (ChREBP): ROLE OF AMP AS AN ALLOSTERIC INHIBITOR.
abstract: The carbohydrate-response element-binding protein (ChREBP) is a glucose-responsive transcription factor that plays an essential role in converting excess carbohydrate to fat storage in the liver. In response to glucose levels, ChREBP is regulated by nuclear/cytosol trafficking via interaction with 14-3-3 proteins, CRM-1 (exportin-1 or XPO-1), or importins. Nuclear localization of ChREBP was rapidly inhibited when incubated in branched-chain α-ketoacids, saturated and unsaturated fatty acids, or 5-aminoimidazole-4-carboxamide ribonucleotide. Here, we discovered that protein-free extracts of high fat-fed livers contained, in addition to ketone bodies, a new metabolite, identified as AMP, which specifically activates the interaction between ChREBP and 14-3-3. The crystal structure showed that AMP binds directly to the N terminus of ChREBP-α2 helix. Our results suggest that AMP inhibits the nuclear localization of ChREBP through an allosteric activation of ChREBP/14-3-3 interactions and not by activation of AMPK. AMP and ketone bodies together can therefore inhibit lipogenesis by restricting localization of ChREBP to the cytoplasm during periods of ketosis.
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Shogo
authors.initials: S
authors.lastname: Sato
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Hunmin
authors.initials: H
authors.lastname: Jung
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakagawa
authors.affiliation: the National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland 20892-8115, and.
authors.forename: Robert
authors.initials: R
authors.lastname: Pawlosky
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Takeshima
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Wan-Ru
authors.initials: WR
authors.lastname: Lee
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Sakiyama
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Sunil
authors.initials: S
authors.lastname: Laxman
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: R Max
authors.initials: RM
authors.lastname: Wynn
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Tu
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: John B
authors.initials: JB
authors.lastname: MacMillan
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390.
authors.forename: Jef K
authors.initials: JK
authors.lastname: De Brabander
authors.affiliation: the National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland 20892-8115, and.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Veech
authors.affiliation: From the Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390, the Dallas Veterans Affairs Medical Center, Dallas, Texas 75216 kuyeda@utsouthwestern.edu.
authors.forename: Kosaku
authors.initials: K
authors.lastname: Uyeda
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism; D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism*; D000494:Allosteric Regulation; D000818:Animals; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D018360:Crystallography, X-Ray; D059305:Diet, High-Fat; D019422:Dietary Sucrose / Q000008:administration & dosage; D022781:Hepatocytes / Q000378:metabolism; D028884:Karyopherins / Q000378:metabolism; D007657:Ketone Bodies / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D051381:Rats; D017207:Rats, Sprague-Dawley; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D000097606:Exportin 1 Protein
chemical_list: D048948:14-3-3 Proteins; D055372:AMP-Activated Protein Kinases; D000249:Adenosine Monophosphate; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D019422:Dietary Sucrose; D028884:Karyopherins; D007657:Ketone Bodies; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C442825:Mlxipl protein, rat
keywords: AMP; ChREBP; allosteric regulation; glucose metabolism; glucose sensing; hepatocyte; ketogenesis; lipogenesis; nuclear localization