Global phosphoproteomics identifies a major role for AKT and 14-3-3 in regulating EDC3

TY - JOUR

T1 - Global phosphoproteomics identifies a major role for AKT and 14-3-3 in regulating EDC3

AU - Larance, Mark

AU - Rowland, Alexander F.

AU - Hoehn, Kyle L.

AU - Humphreys, David T.

AU - Preiss, Thomas

AU - Guilhaus, Michael

AU - James, David E.

PY - 2010/4

Y1 - 2010/4

N2 - Insulin plays an essential role in metabolic homeostasis in mammals, and many of the underlying biochemical pathways are regulated via the canonical phosphatidylinositol 3-kinase/AKT pathway. To identify novel metabolic actions of insulin, we conducted a quantitative proteomics analysis of insulin-regulated 14-3-3-binding proteins in muscle cells. These studies revealed a novel role for insulin in the post-transcriptional regulation of mRNA expression. EDC3, a component of the mRNA decay and translation repression pathway associated with mRNA processing bodies, was shown to be phosphorylated by AKT downstream of insulin signaling. The major insulin-regulated site was mapped to Ser-161, and phosphorylation at this site led to increased 14-3-3 binding. Functional studies indicated that induction of 14-3-3 binding to EDC3 causes morphological changes in processing body structures, inhibition of microRNA-mediated mRNA post-transcriptional regulation, and alterations in the protein- protein interactions of EDC3. These data highlight an important new arm of the insulin signaling cascade in the regulation of mRNA utilization.

AB - Insulin plays an essential role in metabolic homeostasis in mammals, and many of the underlying biochemical pathways are regulated via the canonical phosphatidylinositol 3-kinase/AKT pathway. To identify novel metabolic actions of insulin, we conducted a quantitative proteomics analysis of insulin-regulated 14-3-3-binding proteins in muscle cells. These studies revealed a novel role for insulin in the post-transcriptional regulation of mRNA expression. EDC3, a component of the mRNA decay and translation repression pathway associated with mRNA processing bodies, was shown to be phosphorylated by AKT downstream of insulin signaling. The major insulin-regulated site was mapped to Ser-161, and phosphorylation at this site led to increased 14-3-3 binding. Functional studies indicated that induction of 14-3-3 binding to EDC3 causes morphological changes in processing body structures, inhibition of microRNA-mediated mRNA post-transcriptional regulation, and alterations in the protein- protein interactions of EDC3. These data highlight an important new arm of the insulin signaling cascade in the regulation of mRNA utilization.

KW - Animals

KW - 14-3-3 Proteins

KW - MicroRNAs

KW - Cricetulus

KW - Humans

KW - Ribonucleoproteins, Small Nuclear

KW - RNA Stability

KW - Mice

KW - NIH 3T3 Cells

KW - Binding Sites

KW - Oncogene Protein v-akt

KW - Protein Interaction Mapping

KW - Phosphorylation

KW - Phosphoproteins

KW - Cells, Cultured

KW - Proteomics

KW - CHO Cells

KW - Gene Expression Regulation

KW - RNA Interference

KW - Cricetinae

U2 - 10.1074/mcp.M900435-MCP200

DO - 10.1074/mcp.M900435-MCP200

M3 - Article

C2 - 20051463

SN - 1535-9484

VL - 9

SP - 682

EP - 694

JO - Molecular & Cellular Proteomics

JF - Molecular & Cellular Proteomics

IS - 4

ER -