The large intracytoplasmic loop of the glucose transporter GLUT2 is involved in glucose signaling in hepatic cells.
The hypothesis that the glucose transporter GLUT2 can function as a protein mediating transcriptional glucose signaling was addressed. To divert the putative interacting proteins from a glucose signaling pathway, two intracytoplasmic domains of GLUT2, the C terminus and the large loop located between transmembrane domains 6 and 7, were transfected into mhAT3F hepatoma cells. Glucose-induced accumulation of two hepatic gene mRNAs (GLUT2 and L-pyruvate kinase) was specifically inhibited in cells transfected with the GLUT2 loop and not with the GLUT2 C terminus. The dual effects of glucose were dissociated in cells expressing the GLUT2 loop; in fact a normal glucose metabolism into glycogen occurred concomitantly with the inhibition of the glucose-induced transcription. This inhibition by the GLUT2 loop could be due to competitive binding of a protein that normally interacts with endogenous GLUT2. In addition, the GLUT2 loop, tagged with green fluorescent protein (GFP), was located within the nucleus, whereas the GFP and GFP-GLUT2 C-terminal proteins remained in the cytoplasm. In living cells, a fraction (50%) of the expressed GFP-GLUT2 loop translocated rapidly from the cytoplasm to the nucleus in response to high glucose concentration and conversely in the absence of glucose. We conclude that, via protein interactions with its large loop, GLUT2 may transduce a glucose signal from the plasma membrane to the nucleus.
Guillemain
G
G
Endocrinologie Métabolisme et Développement, CNRS - UPR1524, 92190 Meudon, France. leturque@infobiogen.fr.
Loizeau
M
M
Pinçon-Raymond
M
M
Girard
J
J
Leturque
A
A
D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000235:genetics; D002250:Carbon Radioisotopes; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D005632:Fructose / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D051273:Glucose Transporter Type 2; D049452:Green Fluorescent Proteins; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D008112:Liver Glycogen / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014994:Xylose / Q000494:pharmacology
D002250:Carbon Radioisotopes; D051273:Glucose Transporter Type 2; D008112:Liver Glycogen; D008164:Luminescent Proteins; D009004:Monosaccharide Transport Proteins; D010446:Peptide Fragments; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005632:Fructose; D014994:Xylose; D005947:Glucose