Control of liver glycogen synthase activity and intracellular distribution by phosphorylation.
Eukaryotic glycogen synthase activity is regulated by reversible phosphorylation at multiple sites. Of the two GS isoforms found in mammals, the muscle enzyme (muscle glycogen synthase) has received more attention and the relative importance of every known phosphorylation site in the control of its activity and intracellular distribution has been previously addressed. We have analyzed the impact of the dephosphorylation at the homologous sites of the glycogen synthase liver (LGS) isoform. Serine residues at these sites were replaced by non-phosphorylatable alanine residues, singly or in pairs, and the resultant LGS variants were expressed in cultured cells using adenoviral vectors. The sole mutation at site 2 (Ser7) yielded an enzyme that was almost fully active and able to induce glycogen deposition in primary hepatocytes incubated in the absence of glucose and in FTO2B cells, a cell line that does not normally synthesize glycogen. Mutation at site 2 was also sufficient to trigger the aggregation and translocation of LGS from the cytoplasm to the hepatocyte cell cortex in the absence of glucose. However, this redistribution was not observed in hepatocytes incubated without glucose when an additional mutation (E509A), which renders the enzyme inactive, was introduced. This result suggests that LGS translocation is strictly dependent on glycogen synthesis.
Ros
Susana
S
Institute for Research in Biomedicine IRB Barcelona and CIBER de Diabetes y Enfermedades Metabólicas CIBERDEM, Department of Biochemistry and Molecular Biology, University of Barcelona, Barcelona, Spain.
García-Rocha
Mar
M
Domínguez
Jorge
J
Ferrer
Juan C
JC
Guinovart
Joan J
JJ
D000256:Adenoviridae; D019943:Amino Acid Substitution; D000818:Animals; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D005947:Glucose / Q000378:metabolism*; D006003:Glycogen / Q000096:biosynthesis* / Q000235:genetics; D006006:Glycogen Synthase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D020125:Mutation, Missense; D009928:Organ Specificity / Q000502:physiology; D010766:Phosphorylation / Q000235:genetics; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar
D007527:Isoenzymes; D006003:Glycogen; D006006:Glycogen Synthase; D005947:Glucose