caa a22 4500 475771133 CHVBK 20180406123617.0 cr unu---uuuuu 170329e20000601xx s 000 0 eng 10.1007/s001250051369 doi (NATIONALLICENCE)springer-10.1007/s001250051369 Adenovirus-mediated expression of a naturally occurring Asp905Tyr variant of the glycogen-associated regulatory subunit of protein phosphatase-1 in L6 myotubes [Elektronische Daten] [S. K. Rasmussen, L. Hansen, E. U. Frevert, P. T. W. Cohen, B. B. Kahn, O. Pedersen] Aims/hypothesis. The glycogen-associated protein phosphatase-1 (PP1G) is thought to play an important part in the regulation of skeletal muscle glycogen content. We have previously identified an Asp905Tyr polymorphism of the glycogen-associated regulatory subunit of the protein phosphatase 1 (PPP1R3) gene which among healthy subjects was associated with decreased insulin stimulated non-oxidative glucose metabolism, i. e. primary glycogen synthesis. In this study, the functional effect of the polymorphism was examined in vitro.¶Methods. Wild type (PPP1R3-Asp905) and mutant (PPP1R3-Tyr905) PPP1R3 were expressed in L6 myotubes using adenovirus-mediated gene transfer. Basal and insulin-stimulated glucose uptake and glycogen synthesis were measured. Furthermore, the sensitivity of glycogen synthesis to a cyclic AMP agonist was measured.¶Results. Compared with green fluorescent protein-transduced myotubes and non-transduced myotubes, overexpression of PPP1R3-Asp905 and PPP1R3-Tyr905 increased both basal and insulin-stimulated glycogen synthesis approximately twofold. Treatment of both non-transduced and PPP1R3-transduced L6 myotubes with a cAMP agonist decreased both basal and insulin-stimulated glycogen synthesis by about 40 %. Overexpression of PPP1R3 did not affect either basal or insulin-stimulated 2-deoxy-d-glucose uptake compared with green fluorescent protein-transduced cells.¶Conclusion/interpretation. Results obtained from L6 myotubes transduced with PPP1R3-Asp905 or PPP1R3-Tyr905 showed no statistically significant difference. Therefore, the Asp905Tyr variant alone is unlikely to account for the decreased insulin stimulated non-oxidative glucose metabolism observed in the human study reported previously. [Diabetologia (2000) 43: 718-722] Springer-Verlag Berlin Heidelberg, 2000 KeywordsPPP1R3, PP1G, codon 905 polymorphism, Type II diabetes, glycogen synthesis, non-oxidative glucose metabolism, L6 myotubes, adenovirus nationallicence Rasmussen S. K. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut Hansen L. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut Frevert E. U. Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA, US aut Cohen P. T. W. Medical Research Council Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, UK, GB aut Kahn B. B. Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA, US aut Pedersen O. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut https://doi.org/10.1007/s001250051369 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001250051369 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rasmussen S. K. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut NATIONALLICENCE 700 1- Hansen L. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut NATIONALLICENCE 700 1- Frevert E. U. Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA, US aut NATIONALLICENCE 700 1- Cohen P. T. W. Medical Research Council Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, UK, GB aut NATIONALLICENCE 700 1- Kahn B. B. Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA, US aut NATIONALLICENCE 700 1- Pedersen O. Steno Diabetes Center and Hagedorn Research Institute, Gentofte, Denmark, DK aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer