Fructose modulates GLUT5 mRNA stability in differentiated Caco-2 cells: role of cAMP-signalling pathway and PABP (polyadenylated-binding protein)-interacting protein (Paip) 2.
In intestinal cells, levels of the fructose transporter GLUT5 are increased by glucose and to a greater extent by fructose. We investigated the mechanism by which fructose increases GLUT5 expression. In Caco-2 cells, fructose and glucose increased activity of the -2500/+41 GLUT5 promoter to the same extent. cAMP also activated the GLUT5 promoter. However, if a protein kinase A inhibitor was used to block cAMP signalling, extensive GLUT5 mRNA degradation was observed, with no change in basal transcription levels demonstrating the involvement of cAMP in GLUT5 mRNA stability. Indeed, the half-life of GLUT5 mRNA was correlated ( R2=0.9913) with cellular cAMP levels. Fructose increased cAMP concentration more than glucose, accounting for the stronger effect of fructose when compared with that of glucose on GLUT5 production. We identified several complexes between GLUT5 3'-UTR RNA (where UTR stands for untranslated region) and cytosolic proteins that might participate in mRNA processing. Strong binding of a 140 kDa complex I was observed in sugar-deprived cells, with levels of binding lower in the presence of fructose and glucose by factors of 12 and 6 respectively. This may account for differences in the effects of fructose and glucose. In contrast, the amounts of two complexes of 96 and 48 kDa increased equally after stimulation with either glucose or fructose. Finally, PABP (polyadenylated-binding protein)-interacting protein 2, a destabilizing partner of PABP, was identified as a component of GLUT5 3'-UTR RNA-protein complexes. We conclude that the post-transcriptional regulation of GLUT5 by fructose involves increases in mRNA stability mediated by the cAMP pathway and Paip2 (PABP-interacting protein 2) binding.
Gouyon
Florence
F
Institut National de la Santé et de la Recherche Médicale U505, Université Pierre et Marie Curie, 15 rue de l'Ecole de Médecine, 75006 Paris, France.
Onesto
Cercina
C
Dalet
Veronique
V
Pages
Gilles
G
Leturque
Armelle
A
Brot-Laroche
Edith
E
D020413:3' Untranslated Regions / Q000378:metabolism; D018938:Caco-2 Cells; D050260:Carbohydrate Metabolism; D002454:Cell Differentiation; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D005632:Fructose / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology; D051296:Glucose Transporter Type 5; D006801:Humans; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D015290:Second Messenger Systems; D015533:Transcriptional Activation
D020413:3' Untranslated Regions; D051296:Glucose Transporter Type 5; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D005632:Fructose; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D005947:Glucose