A Mammalian Homologue of GCN2 Protein Kinase Important for Translational Control by Phosphorylation of Eukaryotic Initiation Factor-2

Corresponding author: Ronald C. Wek, Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Van Nuys Medical Science Bldg., Rm. 4067, 635 Barnhill Dr., Indianapolis, IN 46202-5122., rwek{at}iupui.edu (E-mail)

Communicating editor: M. JOHNSTON

A family of protein kinases regulates translation in response to different cellular stresses by phosphorylation of the subunit of eukaryotic initiation factor-2 (eIF-2). In yeast, an eIF-2 kinase, GCN2, functions in translational control in response to amino acid starvation. It is thought that uncharged tRNA that accumulates during amino acid limitation binds to sequences in GCN2 homologous to histidyl-tRNA synthetase (HisRS) enzymes, leading to enhanced kinase catalytic activity. Given that starvation for amino acids also stimulates phosphorylation of eIF-2 in mammalian cells, we searched for and identified a GCN2 homologue in mice. We cloned three different cDNAs encoding mouse GCN2 isoforms, derived from a single gene, that vary in their amino-terminal sequences. Like their yeast counterpart, the mouse GCN2 isoforms contain HisRS-related sequences juxtaposed to the kinase catalytic domain. While GCN2 mRNA was found in all mouse tissues examined, the isoforms appear to be differentially expressed. Mouse GCN2 expressed in yeast was found to inhibit growth by hyperphosphorylation of eIF-2, requiring both the kinase catalytic domain and the HisRS-related sequences. Additionally, lysates prepared from yeast expressing mGCN2 were found to phosphorylate recombinant eIF-2 substrate. Mouse GCN2 activity in both the in vivo and in vitro assays required the presence of serine-51, the known regulatory phosphorylation site in eIF-2. Together, our studies identify a new mammalian eIF-2 kinase, GCN2, that can mediate translational control.

This article has been cited by other articles:

				B. M. Tsoi, A. G. Beckhouse, C. L. Gelling, M. J. Raftery, J. Chiu, A. M. Tsoi, L. Lauterbach, P. J. Rogers, V. J. Higgins, and I. W. Dawes Essential Role of One-carbon Metabolism and Gcn4p and Bas1p Transcriptional Regulators during Adaptation to Anaerobic Growth of Saccharomyces cerevisiae J. Biol. Chem., April 24, 2009; 284(17): 11205 - 11215. [Abstract] [Full Text] [PDF]

				S. Carra, J. F. Brunsting, H. Lambert, J. Landry, and H. H. Kampinga HspB8 Participates in Protein Quality Control by a Non-chaperone-like Mechanism That Requires eIF2{alpha} Phosphorylation J. Biol. Chem., February 27, 2009; 284(9): 5523 - 5532. [Abstract] [Full Text] [PDF]

				N. Su and M. S. Kilberg C/EBP Homology Protein (CHOP) Interacts with Activating Transcription Factor 4 (ATF4) and Negatively Regulates the Stress-dependent Induction of the Asparagine Synthetase Gene J. Biol. Chem., December 12, 2008; 283(50): 35106 - 35117. [Abstract] [Full Text] [PDF]

				X. Wang, B. D. Fonseca, H. Tang, R. Liu, A. Elia, M. J. Clemens, U.-A. Bommer, and C. G. Proud Re-evaluating the Roles of Proposed Modulators of Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling J. Biol. Chem., November 7, 2008; 283(45): 30482 - 30492. [Abstract] [Full Text] [PDF]

				A. Gjymishka, S. S. Palii, J. Shan, and M. S. Kilberg Despite Increased ATF4 Binding at the C/EBP-ATF Composite Site following Activation of the Unfolded Protein Response, System A Transporter 2 (SNAT2) Transcription Activity Is Repressed in HepG2 Cells J. Biol. Chem., October 10, 2008; 283(41): 27736 - 27747. [Abstract] [Full Text] [PDF]

				D. Dardevet, S. R Kimball, L. S Jefferson, A. D Cherrington, D. Remond, C. A DiCostanzo, and M. C. Moore Portal infusion of amino acids is more efficient than peripheral infusion in stimulating liver protein synthesis at the same hepatic amino acid load in dogs Am. J. Clinical Nutrition, October 1, 2008; 88(4): 986 - 996. [Abstract] [Full Text] [PDF]

				N. Niknejad, M. Morley, and J. Dimitroulakos Activation of the Integrated Stress Response Regulates Lovastatin-induced Apoptosis J. Biol. Chem., October 12, 2007; 282(41): 29748 - 29756. [Abstract] [Full Text] [PDF]

				R. Chaturvedi, M. Asim, N. D. Lewis, H. M. S. Algood, T. L. Cover, P. Y. Kim, and K. T. Wilson L-Arginine Availability Regulates Inducible Nitric Oxide Synthase-Dependent Host Defense against Helicobacter pylori Infect. Immun., September 1, 2007; 75(9): 4305 - 4315. [Abstract] [Full Text] [PDF]

				A. M. Silva, D. Wang, A. A. Komar, B. A. Castilho, and B. R. G. Williams Salicylates Trigger Protein Synthesis Inhibition in a Protein Kinase R-like Endoplasmic Reticulum Kinase-dependent Manner J. Biol. Chem., April 6, 2007; 282(14): 10164 - 10171. [Abstract] [Full Text] [PDF]

				A. K. Padyana, H. Qiu, A. Roll-Mecak, A. G. Hinnebusch, and S. K. Burley Structural Basis for Autoinhibition and Mutational Activation of Eukaryotic Initiation Factor 2{alpha} Protein Kinase GCN2 J. Biol. Chem., August 12, 2005; 280(32): 29289 - 29299. [Abstract] [Full Text] [PDF]

				C. M. Pereira, E. Sattlegger, H.-Y. Jiang, B. M. Longo, C. B. Jaqueta, A. G. Hinnebusch, R. C. Wek, L. E. A. M. Mello, and B. A. Castilho IMPACT, a Protein Preferentially Expressed in the Mouse Brain, Binds GCN1 and Inhibits GCN2 Activation J. Biol. Chem., August 5, 2005; 280(31): 28316 - 28323. [Abstract] [Full Text] [PDF]

				S. Hao, J. W. Sharp, C. M. Ross-Inta, B. J. McDaniel, T. G. Anthony, R. C. Wek, D. R. Cavener, B. C. McGrath, J. B. Rudell, T. J. Koehnle, et al. Uncharged tRNA and Sensing of Amino Acid Deficiency in Mammalian Piriform Cortex Science, March 18, 2005; 307(5716): 1776 - 1778. [Abstract] [Full Text] [PDF]

				K. Inoki, H. Ouyang, Y. Li, and K.-L. Guan Signaling by Target of Rapamycin Proteins in Cell Growth Control Microbiol. Mol. Biol. Rev., March 1, 2005; 69(1): 79 - 100. [Abstract] [Full Text] [PDF]

				M. Boyce, K. F. Bryant, C. Jousse, K. Long, H. P. Harding, D. Scheuner, R. J. Kaufman, D. Ma, D. M. Coen, D. Ron, et al. A Selective Inhibitor of eIF2{alpha} Dephosphorylation Protects Cells from ER Stress Science, February 11, 2005; 307(5711): 935 - 939. [Abstract] [Full Text] [PDF]

				E. Gomez, M. L. Powell, I. C. Greenman, and T. P. Herbert Glucose-stimulated Protein Synthesis in Pancreatic {beta}-Cells Parallels an Increase in the Availability of the Translational Ternary Complex (eIF2-GTP{middle dot}Met-tRNAi) and the Dephosphorylation of eIF2{alpha} J. Biol. Chem., December 24, 2004; 279(52): 53937 - 53946. [Abstract] [Full Text] [PDF]

				K. Zhan, J. Narasimhan, and R. C. Wek Differential Activation of eIF2 Kinases in Response to Cellular Stresses in Schizosaccharomyces pombe Genetics, December 1, 2004; 168(4): 1867 - 1875. [Abstract] [Full Text] [PDF]

				T. G. Anthony, B. J. McDaniel, R. L. Byerley, B. C. McGrath, D. R. Cavener, M. A. McNurlan, and R. C. Wek Preservation of Liver Protein Synthesis during Dietary Leucine Deprivation Occurs at the Expense of Skeletal Muscle Mass in Mice Deleted for eIF2 Kinase GCN2 J. Biol. Chem., August 27, 2004; 279(35): 36553 - 36561. [Abstract] [Full Text] [PDF]

				S. Wu, M. Tan, Y. Hu, J.-L. Wang, D. Scheuner, and R. J. Kaufman Ultraviolet Light Activates NF{kappa}B through Translational Inhibition of I{kappa}B{alpha} Synthesis J. Biol. Chem., August 13, 2004; 279(33): 34898 - 34902. [Abstract] [Full Text] [PDF]

				P. Anderson, K. Phillips, G. Stoecklin, and N. Kedersha Post-transcriptional regulation of proinflammatory proteins J. Leukoc. Biol., July 1, 2004; 76(1): 42 - 47. [Abstract] [Full Text] [PDF]

				E. Klann, M. D. Antion, J. L. Banko, and L. Hou Synaptic Plasticity and Translation Initiation Learn. Mem., July 1, 2004; 11(4): 365 - 372. [Abstract] [Full Text] [PDF]

				D. W. Gietzen, C. M. Ross, S. Hao, and J. W. Sharp Phosphorylation of eIF2{alpha} Is Involved in the Signaling of Indispensable Amino Acid Deficiency in the Anterior Piriform Cortex of the Brain in Rats J. Nutr., April 1, 2004; 134(4): 717 - 723. [Abstract] [Full Text] [PDF]

				H.-Y. Jiang, S. A. Wek, B. C. McGrath, D. Lu, T. Hai, H. P. Harding, X. Wang, D. Ron, D. R. Cavener, and R. C. Wek Activating Transcription Factor 3 Is Integral to the Eukaryotic Initiation Factor 2 Kinase Stress Response Mol. Cell. Biol., February 1, 2004; 24(3): 1365 - 1377. [Abstract] [Full Text] [PDF]

				J. Fernandez, A. B. Lopez, C. Wang, R. Mishra, L. Zhou, I. Yaman, M. D. Snider, and M. Hatzolgou Transcriptional Control of the Arginine/Lysine Transporter, Cat-1, by Physiological Stress J. Biol. Chem., December 12, 2003; 278(50): 50000 - 50009. [Abstract] [Full Text] [PDF]

				S. B. Cullinan, D. Zhang, M. Hannink, E. Arvisais, R. J. Kaufman, and J. A. Diehl Nrf2 Is a Direct PERK Substrate and Effector of PERK-Dependent Cell Survival Mol. Cell. Biol., October 15, 2003; 23(20): 7198 - 7209. [Abstract] [Full Text] [PDF]

				H.-Y. Jiang, S. A. Wek, B. C. McGrath, D. Scheuner, R. J. Kaufman, D. R. Cavener, and R. C. Wek Phosphorylation of the {alpha} Subunit of Eukaryotic Initiation Factor 2 Is Required for Activation of NF-{kappa}B in Response to Diverse Cellular Stresses Mol. Cell. Biol., August 15, 2003; 23(16): 5651 - 5663. [Abstract] [Full Text] [PDF]

				L. S. Jefferson and S. R. Kimball Amino Acids as Regulators of Gene Expression at the Level of mRNA Translation J. Nutr., June 1, 2003; 133(6): 2046S - 2051. [Abstract] [Full Text] [PDF]

				R. van Huizen, J. L. Martindale, M. Gorospe, and N. J. Holbrook P58IPK, a Novel Endoplasmic Reticulum Stress-inducible Protein and Potential Negative Regulator of eIF2alpha Signaling J. Biol. Chem., April 25, 2003; 278(18): 15558 - 15564. [Abstract] [Full Text] [PDF]

				Y. Shi, S. I. Taylor, S.-L. Tan, and N. Sonenberg When Translation Meets Metabolism: Multiple Links to Diabetes Endocr. Rev., February 1, 2003; 24(1): 91 - 101. [Abstract] [Full Text] [PDF]

				P. Zhang, B. C. McGrath, J. Reinert, D. S. Olsen, L. Lei, S. Gill, S. A. Wek, K. M. Vattem, R. C. Wek, S. R. Kimball, et al. The GCN2 eIF2{alpha} Kinase Is Required for Adaptation to Amino Acid Deprivation in Mice Mol. Cell. Biol., October 1, 2002; 22(19): 6681 - 6688. [Abstract] [Full Text] [PDF]

				P. J. Bain, R. LeBlanc-Chaffin, H. Chen, S. S. Palii, K. M. Leach, and M. S. Kilberg The Mechanism for Transcriptional Activation of the Human ATA2 Transporter Gene by Amino Acid Deprivation is Different than That for Asparagine Synthetase J. Nutr., October 1, 2002; 132(10): 3023 - 3029. [Abstract] [Full Text] [PDF]

				M. Garcia-Barrio, J. Dong, V. A. Cherkasova, X. Zhang, F. Zhang, S. Ufano, R. Lai, J. Qin, and A. G. Hinnebusch Serine 577 Is Phosphorylated and Negatively Affects the tRNA Binding and eIF2alpha Kinase Activities of GCN2 J. Biol. Chem., August 16, 2002; 277(34): 30675 - 30683. [Abstract] [Full Text] [PDF]

				D. Y. Wu, D. C. Tkachuck, R. S. Roberson, and W. H. Schubach The Human SNF5/INI1 Protein Facilitates the Function of the Growth Arrest and DNA Damage-inducible Protein (GADD34) and Modulates GADD34-bound Protein Phosphatase-1 Activity J. Biol. Chem., July 26, 2002; 277(31): 27706 - 27715. [Abstract] [Full Text] [PDF]

				M. S. Kilberg and I. P. Barbosa-Tessmann Genomic Sequences Necessary for Transcriptional Activation by Amino Acid Deprivation of Mammalian Cells J. Nutr., July 1, 2002; 132(7): 1801 - 1804. [Abstract] [Full Text] [PDF]

				P. Zhang, B. McGrath, S.'a. Li, A. Frank, F. Zambito, J. Reinert, M. Gannon, K. Ma, K. McNaughton, and D. R. Cavener The PERK Eukaryotic Initiation Factor 2{alpha} Kinase Is Required for the Development of the Skeletal System, Postnatal Growth, and the Function and Viability of the Pancreas Mol. Cell. Biol., June 1, 2002; 22(11): 3864 - 3874. [Abstract] [Full Text] [PDF]

				K. Ma, K. M. Vattem, and R. C. Wek Dimerization and Release of Molecular Chaperone Inhibition Facilitate Activation of Eukaryotic Initiation Factor-2 Kinase in Response to Endoplasmic Reticulum Stress J. Biol. Chem., May 17, 2002; 277(21): 18728 - 18735. [Abstract] [Full Text] [PDF]

				S. R. Kimball Regulation of Global and Specific mRNA Translation by Amino Acids J. Nutr., May 1, 2002; 132(5): 883 - 886. [Abstract] [Full Text] [PDF]

				S. Kebache, D. Zuo, E. Chevet, and L. Larose Modulation of protein translation by Nck-1 PNAS, April 16, 2002; 99(8): 5406 - 5411. [Abstract] [Full Text] [PDF]

				J. Fernandez, B. Bode, A. Koromilas, J. A. Diehl, I. Krukovets, M. D. Snider, and M. Hatzoglou Translation Mediated by the Internal Ribosome Entry Site of the cat-1 mRNA Is Regulated by Glucose Availability in a PERK Kinase-dependent Manner J. Biol. Chem., March 29, 2002; 277(14): 11780 - 11787. [Abstract] [Full Text] [PDF]

				A. G. Hinnebusch and K. Natarajan Gcn4p, a Master Regulator of Gene Expression, Is Controlled at Multiple Levels by Diverse Signals of Starvation and Stress Eukaryot. Cell, February 1, 2002; 1(1): 22 - 32. [Full Text] [PDF]

				F. Siu, C. Chen, C. Zhong, and M. S. Kilberg CCAAT/Enhancer-binding Protein-beta Is a Mediator of the Nutrient-sensing Response Pathway That Activates the Human Asparagine Synthetase Gene J. Biol. Chem., December 14, 2001; 276(51): 48100 - 48107. [Abstract] [Full Text] [PDF]

				L. Lu, A.-P. Han, and J.-J. Chen Translation Initiation Control by Heme-Regulated Eukaryotic Initiation Factor 2alpha Kinase in Erythroid Cells under Cytoplasmic Stresses Mol. Cell. Biol., December 1, 2001; 21(23): 7971 - 7980. [Abstract] [Full Text] [PDF]

				T. G. Anthony, A. K. Reiter, J. C. Anthony, S. R. Kimball, and L. S. Jefferson Deficiency of dietary EAA preferentially inhibits mRNA translation of ribosomal proteins in liver of meal-fed rats Am J Physiol Endocrinol Metab, September 1, 2001; 281(3): E430 - E439. [Abstract] [Full Text] [PDF]

				L. S. Jefferson and S. R. Kimball Amino Acid Regulation of Gene Expression J. Nutr., September 1, 2001; 131(9): 2460S - 2466. [Abstract] [Full Text] [PDF]

				I. Novoa, H. Zeng, H. P. Harding, and D. Ron Feedback Inhibition of the Unfolded Protein Response by GADD34-mediated Dephosphorylation of eIF2{alpha} J. Cell Biol., May 21, 2001; 153(5): 1011 - 1022. [Abstract] [Full Text] [PDF]

				F. L. Erickson, J. Nika, S. Rippel, and E. M. Hannig Minimum Requirements for the Function of Eukaryotic Translation Initiation Factor 2 Genetics, May 1, 2001; 158(1): 123 - 132. [Abstract] [Full Text]

				A. J. Entingh, B. K. Law, and H. L. Moses Induction of the C/EBP Homologous Protein (CHOP) by Amino Acid Deprivation Requires Insulin-Like Growth Factor I, Phosphatidylinositol 3-Kinase, and Mammalian Target of Rapamycin Signaling Endocrinology, January 1, 2001; 142(1): 221 - 228. [Abstract] [Full Text] [PDF]

				J. S. Crosby, P. J. Chefalo, I. Yeh, S. Ying, I. M. London, P. Leboulch, and J.-J. Chen Regulation of hemoglobin synthesis and proliferation of differentiating erythroid cells by heme-regulated eIF-2alpha kinase Blood, November 1, 2000; 96(9): 3241 - 3248. [Abstract] [Full Text] [PDF]

				R. Yang, S. A. Wek, and R. C. Wek Glucose Limitation Induces GCN4 Translation by Activation of Gcn2 Protein Kinase Mol. Cell. Biol., April 15, 2000; 20(8): 2706 - 2717. [Abstract] [Full Text]

				H. Kubota, Y. Sakaki, and T. Ito GI Domain-mediated Association of the Eukaryotic Initiation Factor 2alpha Kinase GCN2 with Its Activator GCN1 Is Required for General Amino Acid Control in Budding Yeast J. Biol. Chem., June 30, 2000; 275(27): 20243 - 20246. [Abstract] [Full Text] [PDF]

				J. K. Tyzack, X. Wang, G. J. Belsham, and C. G. Proud ABC50 Interacts with Eukaryotic Initiation Factor 2 and Associates with the Ribosome in an ATP-dependent Manner J. Biol. Chem., October 27, 2000; 275(44): 34131 - 34139. [Abstract] [Full Text] [PDF]

				J. Nika, W. Yang, G. D. Pavitt, A. G. Hinnebusch, and E. M. Hannig Purification and Kinetic Analysis of eIF2B from Saccharomyces cerevisiae J. Biol. Chem., August 18, 2000; 275(34): 26011 - 26017. [Abstract] [Full Text] [PDF]

				D. D. Williams, G. D. Pavitt, and C. G. Proud Characterization of the Initiation Factor eIF2B and Its Regulation in Drosophila melanogaster J. Biol. Chem., February 2, 2001; 276(6): 3733 - 3742. [Abstract] [Full Text] [PDF]

				E. J. Smith, I. Marie, A. Prakash, A. Garcia-Sastre, and D. E. Levy IRF3 and IRF7 Phosphorylation in Virus-infected Cells Does Not Require Double-stranded RNA-dependent Protein Kinase R or Ikappa B Kinase but Is Blocked by Vaccinia Virus E3L Protein J. Biol. Chem., March 16, 2001; 276(12): 8951 - 8957. [Abstract] [Full Text] [PDF]

				J. Fernandez, I. Yaman, W. C. Merrick, A. Koromilas, R. C. Wek, R. Sood, J. Hensold, and M. Hatzoglou Regulation of Internal Ribosome Entry Site-mediated Translation by Eukaryotic Initiation Factor-2alpha Phosphorylation and Translation of a Small Upstream Open Reading Frame J. Biol. Chem., January 11, 2002; 277(3): 2050 - 2058. [Abstract] [Full Text] [PDF]