title: Identification of a novel AS160 splice variant that regulates GLUT4 translocation and glucose-uptake in rat muscle cells.
abstract: AS160 (AKT substrate of 160 kDa) is an important mediator of GLUT4 (glucose transporter 4) translocation and glucose-uptake in adipocytes and muscle cells. In our study we have identified a novel splice variant of AS160 (variant 2 of AS160, AS160_v2) that lacks exon 11 and 12. The protein is phosphorylated in response to insulin via the PI3K/AKT pathway. Expression of this splice variant in human tissues from different donors was examined with quantitative RT-PCR. Our data reveal a tissue specific distribution pattern of both isoforms with highest overall expression of AS160_v2. To investigate the function of the novel splice variant we established the doxycycline-inducible expression of the protein in a rat myoblast cell line co-expressing GLUT4-myc. In contrast to data reported for the full-length AS160 protein, over expression and activation of transcript variant 2 in this cell line increased GLUT4 translocation and glucose-uptake rates in response to insulin and IGF-1 but not in response to AICAR or metformin. Immunofluorescence based studies indicated a direct association of AS160_v2 with GLUT4 under basal but not under insulin-stimulated conditions. Additionally, over expression of AS160_v2 slightly improved glucose-uptake rates in a model of insulin resistance but was not able to fully prevent induction of insulin resistance. This was accompanied with decreased phosphorylation of AS160_v2 and AKT. Taken together, our data suggest a tissue specific distribution of full-length AS160 and the novel AS160 splice variant (AS160_v2) indicating different functions. In contrast to full-length AS160, transcript variant 2 of AS160 seems to be a novel regulator of glucose transport that positively influences glucose-uptake rates.
authors.affiliation: Sanofi-Aventis Germany GmbH, TD Metabolism, 65926 Frankfurt am Main, Germany. daniela.baus@sanofi-aventis.com
authors.forename: Daniela
authors.initials: D
authors.lastname: Baus
authors.forename: Kathrin
authors.initials: K
authors.lastname: Heermeier
authors.forename: Meltsje
authors.initials: M
authors.lastname: De Hoop
authors.forename: Christiane
authors.initials: C
authors.lastname: Metz-Weidmann
authors.forename: Johann
authors.initials: J
authors.lastname: Gassenhuber
authors.forename: Werner
authors.initials: W
authors.lastname: Dittrich
authors.forename: Stefan
authors.initials: S
authors.lastname: Welte
authors.forename: Norbert
authors.initials: N
authors.lastname: Tennagels
mesh_terms: D017667:Adipocytes / Q000502:physiology; D017398:Alternative Splicing; D000704:Analysis of Variance; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D047569:Laser Scanning Cytometry; D032342:Muscle Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010766:Phosphorylation; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013997:Time Factors; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D020690:GTPase-Activating Proteins; D051275:Glucose Transporter Type 4; D007328:Insulin; D020033:Protein Isoforms; D012333:RNA, Messenger; C518767:TBC1D4 protein, human; D005947:Glucose; D000077191:Wortmannin