| Insulin Signaling and the Control of PHAS-I Phosphorylation | |
| Introduction | p. 1 |
| Mechanism of Translational Repression | p. 1 |
| PHAS Isoforms | p. 3 |
| Phosphorylation Sites in PHAS-I | p. 5 |
| Identification of Sites | p. 6 |
| Influence of Phosphorylation on the Electrophoretic Mobility of PHAS-I | p. 6 |
| Sites Involved in the Control of eIF4E Binding | p. 7 |
| Potential Mechanisms of Ordered Phosphorylation | p. 8 |
| Protein Kinases That Phosphorylate PHAS-I in Vitro | p. 10 |
| mTOR Protein | p. 11 |
| Protein Kinase C | p. 15 |
| Protein Kinase CK2 | p. 16 |
| MAP Kinase | p. 16 |
| Control by Hormones, Nutrients, and cAMP | p. 17 |
| The Insulin Signaling Pathway | p. 18 |
| Insulin Receptor Substrate 1 (IRS-1) | p. 18 |
| Phosphatidyl Inositol 3-OH Kinase (PI 3-kinase) | p. 18 |
| Protein Kinase B | p. 19 |
| mTOR Phosphorylation | p. 20 |
| p. 22 |
| Regulation of PHAS-I by a Nutrient-Sensing Pathway | p. 23 |
| Regulation of PHAS-I Dephosphorylation | p. 25 |
| References | p. 26 |
| Insulin, Phorbol Ester and Serum Regulate the Elongation Phase of Protein Synthesis | |
| Introduction | p. 33 |
| Structure and Function of EF-1 and EF-2 | p. 33 |
| Modifications of EF-1 and EF-2 | p. 37 |
| Regulation of Elongation by Insulin Via Multipotential S6 Kinase and EF-2 Kinase | p. 39 |
| Regulation of Elongation by Phorbol Ester Via Protein Kinase C | p. 42 |
| Regulation of Elongation during the Cell Cycle by Cdc2 | p. 43 |
| Lack of Regulation of Elongation by Protein Kinase Casein Kinase II | p. 44 |
| Conclusions | p. 45 |
| References | p. 45 |
| Regulation of Protein Synthesis by Insulin Through IRS-1 | |
| Introduction | p. 49 |
| Materials and Methods | p. 52 |
| Cell Lines | p. 52 |
| Measurement of Protein and DNA Synthesis | p. 53 |
| MAPK, p70(S6K), PI3K and PKC Activity | p. 53 |
| Preparation of (32)P-Labeled eIF4E and PHAS-I | p. 54 |
| MAPK Depletion | p. 54 |
| Northern Blot Analysis | p. 55 |
| eIF2B and GSK-3 Activity | p. 55 |
| Results | p. 55 |
| Both IR and IRS-1 Are Required for Stimulation of Translation by Insulin in 32D Cells | p. 55 |
| MAPK Activation Is Necessary But Not Sufficient for Insulin-Stimulated Protein Synthesis | p. 56 |
| SHP-2 Attenuates the IRS-1 Signal | p. 57 |
| The Insulin Signal to Protein Synthesis Proceeds Through PI3K | p. 59 |
| The mTOR Branch Downstream of PI3K Stimulates Growth-Regulated Translation | p. 62 |
| The PKC$$ Branch Downstream of PI3K Stimulates General Translation | p. 66 |
| General Protein Synthesis Is Correlated with Inhibition of GSK-3 and Activation of eIF2B | p. 71 |
| Discussion | p. 77 |
| Insulin Receptor and Insulin Receptor Substrate-1 | p. 77 |
| GRB-2/SOS Binding to IRS-1 | p. 77 |
| SHP-2 Binding to IRS-1 | p. 78 |
| PI3K Binding to IRS-1 | p. 79 |
| The Rapamycin-Sensitive Branch Involves PKB and mTOR | p. 79 |
| The Rapamycin-Insensitive Branch Proceeds Through PKC$$ | p. 80 |
| Glycogen Synthase Kinase-3 and eIF2B | p. 82 |
| Protein Synthesis and Cell Proliferation | p. 83 |
| Pathway from Insulin to General and Growth-Related Protein Synthesis | p. 84 |
| References | p. 85 |
| Regulation of Eukaryotic Initiation Factor eIF2B | |
| Function and Structure of eIF2B | p. 95 |
| eIF2B Is a Guanine Nucleotide Exchange Factor | p. 95 |
| eIF2B Is a Heteropentameric Protein | p. 95 |
| eIF2B Is an Important Control Point for Translation Initiation | p. 97 |
| eIF2B Activity Can Be Regulated by the Phosphorylation of eIF2$$ | p. 97 |
| Regulation of eIF2B Activity in Vivo | p. 98 |
| Mechanisms Involved in the "Direct" Regulation of eIF2B Activity | p. 100 |
| eIF2B May Be Regulated Allosterically and by Phosphorylation | p. 100 |
| eIF2Be Is a Substrate For GSK-3 | p. 100 |
| Control of GSK-3 Activity | p. 102 |
| Regulation of Phosphorylation of the GSK-3 Site in eIF2Be | p. 104 |
| The Erk Pathway Can Also Modulate eIF2B Activity | p. 104 |
| Other Phosphorylation Sites in eIF2B | p. 106 |
| Phosphorylation of the Priming Site in eIF2Be | p. 106 |
| Phosphorylation Sites in eIF2Be in Vivo | p. 107 |
| Phosphorylation of eIF2B by Casein Kinases | p. 107 |
| Are Other Subunits of eIF2B Phosphorylated? | p. 108 |
| Other Inputs into the Control of eIF2B | p. 109 |
| Conclusions and Perspectives | p. 110 |
| p. 111 |
| The p70 S6 Kinase Integrates Nutrient and Growth Signals to Control Translational Capacity | |
| Identification of the p70 S6 Kinase | p. 115 |
| Expression and Structure | p. 117 |
| Substrate Specificity and Selection | p. 120 |
| Cellular Function(s) | p. 122 |
| The p70 S6 Kinase Controls Expression of the Translational Apparatus by Regulating Initiation of 5' Terminal Oligopyrimidine Sequence mRNAs | p. 122 |
| The p70 S6 Kinase Coordinates Cell Division with Cell Growth | p. 125 |
| Regulation of the p70 S6 Kinase | p. 127 |
| TOR Regulates Cell Function in Response to the Nutrient Milieu | p. 128 |
| p70 is Regulated by Multisite (Ser/Thr) Phosphorylation | p. 132 |
| RTK Recruitment of Type 1A PI-3 Kinases Activates p70 S6 Kinase | p. 134 |
| The Mechanism of p70 Activation by PI-3 Kinase and the Role of PDK1 | p. 135 |
| Candidate "p70 Thr412 Kinases" | p. 140 |
| PDK1 As a p70 Thr412 Kinase | p. 140 |
| mTOR As a p70 Thr412 Kinase | p. 141 |
| A Novel Set of p70 Thr412 Kinases | p. 145 |
| Conclusion | p. 145 |
| p. 146 |
| Regulation of Translation Initiation by Amino Acids in Eukaryotic Cells | |
| Introduction | p. 155 |
| Pathway of Translation Initiation | p. 155 |
| Regulation by Amino Acids of met-tRNAi Binding to 40 S Ribosomal Subunits | p. 157 |
| Regulation of met-tRNAi Binding in Saccharomyces cerevisiae | p. 157 |
| Regulation of GCN4 mRNA Translation by Amino Acids | p. 157 |
| Roles of eIF2 and eIF2B in Translational Regulation of Gcn4p Expression by Amino Acids | p. 159 |
| Gcn2p Is an eIF2a Kinase That Regulates Gcn4p Expression by Amino Acids | p. 161 |
| Model for the Translational Regulation of Gcn4p Expression by Amino Acids | p. 162 |
| Regulation ofmet-tRNAi Binding in Mammalian Cells | p. 163 |
| Regulation of mRNA Binding to 40 S Ribosomal Subunits by Amino Acids | p. 166 |
| Modulation of 4E-BP1 and S6K1 Phosphorylation by Amino Acids | p. 168 |
| Signaling Pathways for Leucine-Mediated Changes in Translation Initiation | p. 172 |
| Is There Coordinated Regulation by Amino Acids of Translation Initiation and Elongation? | p. 174 |
| Summary | p. 175 |
| References | p. 177 |
| Subject Index | p. 185 |
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