Abstract
Childhood ataxia with central nervous system hypomyelination (CACH), also called vanishing white matter (VWM) leukoencephalopathy, is a fatal genetic disease caused by mutations in eukaryotic initiation factor 2B (eIF2B) genes. The five subunits eIF2B factor is critical for translation initiation under normal conditions and regulates protein synthesis in response to cellular stresses. Primary fibroblasts from CACH/VWM patients and normal individuals were used to measure basal eIF2B activity as well as global protein synthesis and ATF4 induction in response to stress in the endoplasmic reticulum. We show that although the cells expressing mutant eIF2B genes respond normally to stress conditions by reduced global translation rates, they exhibit significantly greater increase in ATF4 induction compared to normal controls despite equal levels of stress and activity of the upstream eIF2α kinase. This heightened stress response observed in primary fibroblasts that suffer from minor loss of basal eIF2B activity may be employed as an initial screening tool for CACH/VWM leukodystrophy.
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Balachandran S, Barber GN (2004) Defective translational control facilitates vesicular stomatitis virus oncolysis. Cancer Cell 5:51–65
Baumann N, Pham-Dinh D (2001) Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev 81:871–927
Brostrom CO, Brostrom MA (1998) Regulation of translational initiation during cellular responses to stress. Prog Nucleic Acid Res Mol Biol 58:79–125
Calfon M, Zeng H, Urano F, Till JH, Hubbard SR, Harding HP, Clark SG, Ron D (2002) IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA. Nature 415:92–96
Clemens MJ (2001) Initiation factor eIF2 alpha phosphorylation in stress responses and apoptosis. Prog Mol Subcell Biol 27:57–89
DeGracia DJ, Kumar R, Owen CR, Krause GS, White BC (2002) Molecular pathways of protein synthesis inhibition during brain reperfusion: implications for neuronal survival or death. J Cereb Blood Flow Metab 22:127–141
Dever TE (2002) Gene-specific regulation by general translation factors. Cell 108:545–556
Fogli A, Dionisi-Vici C, Deodato F, Bartuli A, Boespflug-Tanguy O, Bertini E (2002a) A severe variant of childhood ataxia with central hypomyelination/vanishing white matter leukoencephalopathy related to EIF21B5 mutation. Neurology 59:1966–1968
Fogli A, Wong K, Eymard-Pierre E, Wenger J, Bouffard JP, Goldin E, Black DN, Boespflug-Tanguy O, Schiffmann R (2002b) Cree leukoencephalopathy and CACH/VWM disease are allelic at the EIF2B5 locus. Ann Neurol 52:506–510
Fogli A, Rodriguez D, Eymard-Pierre E, Bouhour F, Labauge P, Meaney BF, Zeesman S, Kaneski CR, Schiffmann R, Boespflug-Tanguy O (2003) Ovarian failure related to eukaryotic initiation factor 2B mutations. Am J Hum Genet 72:1544–1550
Fogli A, Schiffmann R, Hugendubler L, Combes P, Bertini E, Rodriguez D, Kimball SR, Boespflug-Tanguy O (2004) Decreased guanine nucleotide exchange factor activity in eIF2B-mutated patients. Eur J Hum Genet 12:561–566
Fromm M, Berg P (1982) Deletion mapping of DNA regions required for SV40 early region promoter function in vivo. J Mol Appl Genet 1:457–481
Gerlitz G, Jagus R, Elroy-Stein O (2002) Phosphorylation of initiation factor-2 alpha is required for activation of internal translation initiation during cell differentiation. Eur J Biochem 269:2810–2819
Harding HP, Novoa I, Zhang Y, Zeng H, Wek R, Schapira M, Ron D (2000) Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell 6:1099–1108
Harding HP, Calfon M, Urano F, Novoa I, Ron D (2002) Transcriptional and translational control in the Mammalian unfolded protein response. Annu Rev Cell Dev Biol 18:575–599
Hinnebusch AG (2000) Mechanism and regulation of initiator methionyl-tRNA binding to ribosomes. In: Sonenberg N, Hershey JWB, Mathews MB (eds) Translational control of gene expression. Cold Spring Harbor Laboratory Press, New York, pp 185–243
Hinnebusch AG, Natarajan K (2002) Gcn4p, a master regulator of gene expression, is controlled at multiple levels by diverse signals of starvation and stress. Eukaryot Cell 1:22–32
Kaufman RJ (1999) Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev 13:1211–1233
Kaufman RJ (2002) Orchestrating the unfolded protein response in health and disease. J Clin Invest 110:1389–1398
Kimball SR, Everson WV, Myers LM, Jefferson LS (1987) Purification and characterization of eukaryotic initiation factor 2 and a guanine nucleotide exchange factor from rat liver. J Biol Chem 262:2220–2227
Kimball SR, Everson WV, Flaim KE, Jefferson LS (1989) Initiation of protein synthesis in a cell-free system prepared from rat hepatocytes. Am J Physiol 256:C28–C34
van der Knaap MS, Leegwater PA, Konst AA, Visser A, Naidu S, Oudejans CB, Schutgens RB, Pronk JC (2002) Mutations in each of the five subunits of translation initiation factor eIF2B can cause leukoencephalopathy with vanishing white matter. Ann Neurol 51:264–270
Koumenis C, Naczki C, Koritzinsky M, Rastani S, Diehl A, Sonenberg N, Koromilas A, Wouters BG (2002) Regulation of protein synthesis by hypoxia via activation of the endoplasmic reticulum kinase PERK and phosphorylation of the translation initiation factor eIF2alpha. Mol Cell Biol 22:7405–7416
Larocca JN, Rodriguez-Gabin AG (2002) Myelin biogenesis: vesicle transport in oligodendrocytes. Neurochem Res 27:1313–1329
Leegwater PA, Vermeulen G, Konst AA, Naidu S, Mulders J, Visser A, Kersbergen P, Mobach D, Fonds D, van Berkel CG, Lemmers RJ, Frants RR, Oudejans CB, Schutgens RB, Pronk JC, van der Knaap MS (2001) Subunits of the translation initiation factor eIF2B are mutant in leukoencephalopathy with vanishing white matter. Nat Genet 29:383–388
Li W, Wang X, Van Der Knaap MS, Proud CG (2004) Mutations linked to leukoencephalopathy with vanishing white matter impair the function of the eukaryotic initiation factor 2B complex in diverse ways. Mol Cell Biol 24:3295–3306
Lu PD, Harding HP, Ron D (2004) Translation reinitiation at alternative open reading frames regulates gene expression in an integrated stress response. J Cell Biol 167:27–33
Martin de la Vega C, Burda J, Nemethova M, Quevedo C, Alcazar A, Martin ME, Danielisova V, Fando JL, Salinas M (2001a) Possible mechanisms involved in the down-regulation of translation during transient global ischaemia in the rat brain. Biochem J 357:819–826
Martin de la Vega C, Burda J, Salinas M (2001b) Ischemia-induced inhibition of the initiation factor 2alpha phosphatase activity in the rat brain. Neuroreport 12:1021–1025
Paschen W (2003) Endoplasmic reticulum: a primary target in various acute disorders and degenerative diseases of the brain. Cell Calcium 34:365–383
Raine CS (1997) Oligodendrocytes and central nervous system myelin. In: Robertson RLDaDM (ed) Textbook of neuropathology, 3rd edn. Williams& Wilkins, Baltimore, pp 137–164
Richardson JP, Mohammad SS, Pavitt GD (2004) Mutations causing childhood ataxia with central nervous system hypomyelination reduce eukaryotic initiation factor 2B complex formation and activity. Mol Cell Biol 24:2352–2363
Schiffmann R, Moller JR, Trapp BD, Shih HH, Farrer RG, Katz DA, Alger JR, Parker CC, Hauer PE, Kaneski CR et al (1994) Childhood ataxia with diffuse central nervous system hypomyelination. Ann Neurol 35:331–340
Schiffmann R, Tedeschi G, Kinkel RP, Trapp BD, Frank JA, Kaneski CR, Brady RO, Barton NW, Nelson L, Yanovski JA (1997) Leukodystrophy in patients with ovarian dysgenesis. Ann Neurol 41:654–661
Scorsone KA, Panniers R, Rowlands AG, Henshaw EC (1987) Phosphorylation of eukaryotic initiation factor 2 during physiological stresses which affect protein synthesis. J Biol Chem 262:14538–14543
Vallejo M, Ron D, Miller CP, Habener JF (1993) C/ATF, a member of the activating transcription factor family of DNA-binding proteins, dimerizes with CAAT/enhancer-binding proteins and directs their binding to cAMP response elements. Proc Natl Acad Sci USA 90:4679–4483
Van Der Knaap MS, Van Berkel CG, Herms J, Van Coster R, Baethmann M, Naidu S, Boltshauser E, Willemsen MA, Plecko B, Hoffmann GF, Proud CG, Scheper GC, Pronk JC (2003) eIF2B-related disorders: antenatal onset and involvement of multiple organs. Am J Hum Genet 73:1199–1207
Vattem KM, Wek RC (2004) Reinitiation involving upstream ORFs regulates ATF4 mRNA translation in mammalian cells. Proc Natl Acad Sci USA 101:11269–11274
Wang X, Paulin FE, Campbell LE, Gomez E, O’Brien K, Morrice N, Proud CG (2001) Eukaryotic initiation factor 2B: identification of multiple phosphorylation sites in the epsilon-subunit and their functions in vivo. Embo J 20:4349–4359
Wong K, Armstrong RC, Gyure KA, Morrison AL, Rodriguez D, Matalon R, Johnson AB, Wollmann R, Gilbert E, Le TQ, Bradley CA, Crutchfield K, Schiffmann R (2000) Foamy cells with oligodendroglial phenotype in childhood ataxia with diffuse central nervous system hypomyelination syndrome. Acta Neuropathol (Berl) 100:635–646
Acknowledgements
We thank Ehud Goldin for his critical role in establishing the initial phase of the project and for discussions along the way and Lynne Hugendubler for doing the eIF2B assays. This research was supported by The Israel Science Foundation (grant# 439/01-04) to OES;US-Israel Binational Science Foundation (grant# 2003364) to OES and DR, NIH grants DK47119 and ES08681 to DR and NIH grant DK15658 to SRK.
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Kantor, L., Harding, H.P., Ron, D. et al. Heightened stress response in primary fibroblasts expressing mutant eIF2B genes from CACH/VWM leukodystrophy patients. Hum Genet 118, 99–106 (2005). https://doi.org/10.1007/s00439-005-0024-x
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DOI: https://doi.org/10.1007/s00439-005-0024-x