title: An essential role for the Glut1 PDZ-binding motif in growth factor regulation of Glut1 degradation and trafficking.
abstract: Cell surface localization of the Glut (glucose transporter), Glut1, is a cytokine-controlled process essential to support the metabolism and survival of haemopoietic cells. Molecular mechanisms that regulate Glut1 trafficking, however, are not certain. In the present study, we show that a C-terminal PDZ-binding motif in Glut1 is critical to promote maximal cytokine-stimulated Glut1 cell surface localization and prevent Glut1 lysosomal degradation in the absence of growth factor. Disruption of this PDZ-binding sequence through deletion or point mutation sharply decreased surface Glut1 levels and led to rapid targeting of internalized Glut1 to lysosomes for proteolysis, particularly in growth factor-deprived cells. The PDZ-domain protein, GIPC (G(alpha)-interacting protein-interacting protein, C-terminus), bound to Glut1 in part via the Glut1 C-terminal PDZ-binding motif, and we found that GIPC deficiency decreased Glut1 surface levels and glucose uptake. Unlike the Glut1 degradation observed on mutation of the Glut1 PDZ-binding domain, however, GIPC deficiency resulted in accumulation of intracellular Glut1 in a pool distinct from the recycling pathway of the TfR (transferrin receptor). Blockade of Glut1 lysosomal targeting after growth factor withdrawal also led to intracellular accumulation of Glut1, a portion of which could be rapidly restored to the cell surface after growth factor stimulation. These results indicate that the C-terminal PDZ-binding motif of Glut1 plays a key role in growth factor regulation of glucose uptake by both allowing GIPC to promote Glut1 trafficking to the cell surface and protecting intracellular Glut1 from lysosomal degradation after growth factor withdrawal, thus allowing the potential for a rapid return of intracellular Glut1 to the cell surface on restimulation.
authors.affiliation: Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Heather L
authors.initials: HL
authors.lastname: Wieman
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Horn
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Jacobs
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Altman
authors.forename: Sally
authors.initials: S
authors.lastname: Kornbluth
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Rathmell
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism / Q000502:physiology; D000595:Amino Acid Sequence / Q000502:physiology; D000818:Animals; D000954:Antigens, Surface / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D005947:Glucose / Q000378:metabolism; D051272:Glucose Transporter Type 1 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D054731:PDZ Domains / Q000502:physiology*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000954:Antigens, Surface; C492009:GIPC1 protein, human; D051272:Glucose Transporter Type 1; D036341:Intercellular Signaling Peptides and Proteins; D005947:Glucose