Mitochondrial malfunction in vanishing white matter disease: a disease of the cytosolic translation machinery

Mitochondrial malfunction in vanishing white matter disease: a disease of the cytosolic translation machinery

Vanishing white matter (VwM) disease – a disease of the cytosolic translation machinery:VWM is a recessive genetic neurodegenerative disease caused by mutations in any of the five genes encoding the subunits of translation initiation factor 2B (eIF2B) (Leegwater et al., 2001; OMIM 306896).This cure-less disease is rare (rough estimation would be around 1:100,000), but the actual prevalence is thought to be limited by its challenging diagnosis. The disease is characterized by a progressive loss of brain white matter in both hemispheres, causing impairments of neurological functions. Progressive deterioration occurs upon exposure to various physiological stressors, followed eventually by death (Elroy-Stein and Schiffmann, 2015). VWM is in fact a disease of the cytosolictranslational machinery. As such, it illuminates the dependence of glial cells on a no-less-thanperfect translational control.e imperfection due to eIF2B mutations is the sum effects of unbalanced global and specific protein synthesis owing to issues of quantities and coordination, with multiple appearances depending on cellular scenarios. A recent study (Raini et al., 2017) provides evidence that homozygous eIF2B5 R132H mutation in mouse(corresponding to R136H in human) leads to impaired oxidative phosphorylation.is cellular phenotype results from a lack of tight coordination between the cytosolic and mitochondrial translation machineries that synthesize the 83 and 13 electron transfer chain (ETC) subunits, respectively.e abnormal ETC composition, confirmed by unbiased mass spectrometry data and biochemical assays (Gat-Viks et al.,2015; Raini et al., 2017), is one of the deleterious effects of imperfect regulation of gene expression due to partial-lossof-function mutation in eIF2B.

eIF2B is an essential factor required for translation of all mRNAs into proteins within each and every cell type. Due to its unique function (see below), it serves as a master regulator of protein synthesis. Obviously, severe mutations that eliminate eIF2B activity are lethal. However, milder mutations that lead to a tolerated decrease in eIF2B activity are associated with diseased cellular phenotype.e vulnerability of each cell type to eIF2B mutations depends on its functional expertise.

The connection of eIF2B to integrated stress response(ISR) and the differential effect of eIF2B mutation on translation rate of individual mRNAs:eIF2B is the guanine exchange factor (GEF) of eIF2. By recycling the inactive eIF2-GDP back to active eIF2-GTP, eIF2B enables the formation of eIF2-GTP-Met-tRNAiMetternary complex(TC). The cellular concentration of TC dictates the rate of translation initiation. TCs are delivered to 40S ribosomal subunit to generate 43S pre-initiation complexes which enable start codon recognition at each round of translation initiation step. Importantly, the activity of eIF2B is regulated in response to cellular needs. Down-regulation and up-regulation of eIF2B enzymatic activity lead to changes in cellular TC levels, thereby affecting both global and specific mRNA translation. Under stress conditions, one of the four eIF2α kinases (general control nonderepressible 2 (GCN2), proline-rich receptor-like protein kinase (PERK), protein kinase R (PKR) or heme regulated inhibitor (HRI), depending on the stress) is activated and phosphorylates the alpha subunit of eIF2, thereby turning it from a substrate of eIF2B to a competitive inhibitor (Wek et al., 2006).e low eIF2B/eIF2 molar ratio guarantees that even modest eIF2α phosphorylation level will result in an inhibitory effect on eIF2B activity,leading to decreased concentration of 43S pre-initiation complexes.e subsequent differential effect on the translation rate of multiple mRNAs marks the beginning of an expression program termed ISR which also includes feedback dephosphorylation of eIF2α. As predicted, partial-loss-offunction mutations in eIF2B genes are associated with hyper sensitivity to physiological stress (Kantor et al., 2005).is is explained by the fact that eIF2B is constitutively less active,even prior to the stress-induced increase in eIF2α phosphorylation. It is assumed that eIF2B mutations interfere with the tight coordination of the ISR gene expression program.e hyper sensitivity of eIF2B-mutant cells to physiological stress provides an additional reason for the high vulnerabili-ty of glial cells.is is not surprising, because the tasks of all glial cells involve massive production of glycoproteins in the ER, rendering them highly dependent on accurate ER-stress response which obviously requires faultless ISR.

Figure 1 e effect of translation initiation factor 2B (eIF2B) partial-loss-of-function mutation on cellular phenotype.

ISR gene expression program is primarily based on the differential translation of individual mRNAs depending on their 5′-untranslated regions (5′UTR). Specifically, while a decline in the concentration of 43S pre-initiation complexes leads to reduced global translation, it leads (counter intuitively) to enhanced translation of mRNAs harboring regulatory sequences within their 5’UTR (Young and Wek, 2016).Short upstream open reading frames (uORFs) and internal regulatory entry sites (IRES) differentially regulate gene-specific translation in ISR. Key features of uORFs serve to optimize translational control that is essential for regulation of cell fate in response to physiological and environmental stresses. These features guarantee accurate and timely expression of proteins to alleviate the stress and resume global protein synthesis, or alternatively to promote pro-apoptotic signaling cascade. The inborn differential effect of 43S pre-initiation complexes concentration (affected by eIF2B activity) on translation of individual mRNAs, provokes deleterious outcome in cells expressing eIF2B genes with partialloss-of-function mutations. In agreement with this notion,abnormal increased/decreased abundances of multiple proteins were detected by mass-spectroscopy measurements of total proteomes of whole brains and cultured fibroblasts isolated from eIF2B5R132H/R132Hmice. Analysis of the proteome data exposed unbalanced stoichiometry of subunits within large protein complexes. A major influence on the composition of ETC complexes discovered phenotypic outcome related to defective mitochondrial performance and adenosine triphosphate (ATP) production (Raini et al., 2017;Figure 1).

A multilayered effect leads to defective mitochondrial function:e effect of eIF2B mutation on oxidative phosphorylation (oxphos) is a striking demonstration of the intricacy of cellular circuits. It highlights the importance of the functional link between the cytosolic and mitochondrial translation machineries and also illuminates eIF2B as an important coordinator. To generate mitochondria the cell needs to express ～1,000 nuclear- and 13 mitochondrial-encoded protein gene products. Oxidative respiration relies on the utilization of five multi-subunit protein complexes (ETC complex I–IV and ATP-synthase) composed of total of 96 protein gene products, only 13 of which are encoded by the mitochondrial genome. Synthesis and assembly of these protein complexes require tight synchronization of gene expression from the two genomes, including coordinated work of the two separated translation machineries. While eIF2B role is restricted to the cytosol, the mitochondrial translation machinery cannot escape its functional imperfection. The linkage is dictated by the fact that all of the mitochondrial translation machinery components, except mitochondrial ribosomal RNA (mt-rRNA) and mitochondrial transfer RNAs (mt-tRNAs), are nuclear-encoded, thus are translated in the cytosol. Specifically, eIF2B is involved in the synthesis of all mitochondrial ribosomal proteins (MRPs), tRNA synthetases and translation regulatory factors. Altogether,eIF2B partial-loss-of-function mutations can be associated with defective mitochondrial function due to multilayered interconnected events, as illustrated inFigure 1B: (i) unbalanced abundance of ETC subunits that are translated in the cytosol; (ii) deleterious effect on mitochondrial translation efficacy due to unbalanced abundance of its cytosolic-translated components; (iii) enhanced harmful effect on ETC composition due to the two above mentioned problems,thereby leading to defective oxphos. In agreement with this notion is the total proteome data collected from mouse fibroblasts homozygous for eIF2B5 R132H mutation (Raini et al., 2017). It shows a spectrum of positive and negative foldchange values of components of the mitochondrial translation machinery with significant changes in abundance of several MRPs of the 39S mitochondrial ribosome subunit.

More about the immense sensitivity of glial cells to eIF2B mutations:e discovery of defective mitochondrial performance due to partial loss of eIF2B activity was achieved using primary fibroblasts isolated from eIF2B5R132H/R132Hmice.An important note is that fibroblasts do not show any sign of disease in VWM patients. Interestingly, further research revealed that 1.5-fold increase in mitochondrial abundance in eIF2B5-mutant fibroblasts fully compensates for their energy requirements, even without any change in their glycolysis rate, explaining the lack of their involvement in VWM disease. In contrast, 2-fold increase in mitochondrial abundance together with increased glycolysis rate were not sufficient to meet the energy requirement of primary astrocytes isolated from eIF2B5R132H/R132Hmice, explaining their increased sensitivity to eIF2B mutations and their involvement in the disease (Raini et al., 2017).

OES’ work was funded by The Legacy Heritage Bio-Medical Program of the Israel Science Foundation (grant No.1629/13). I thank all OES lab members for their contribution to VWM-related research and Elad Stein for the graphic illustration.

Orna Elroy-Stein＊

Dept of Cell Research and Immunology, George S. Wise Faculty of Life Sciences; Sagol School of Neuroscience, Tel Aviv University,

Israel

＊Correspondence to:Orna Elroy-Stein, Ph.D., ornaes@tauex.tau.ac.il.orcid: 0000-0002-3716-1540 (Orna Elroy-Stein)

Accepted: 2017-10-10

How to cite this article:Elroy-Stein O (2017) Mitochondrial malfunction in vanishing white matter disease: a disease of the cytosolic translation machinery. Neural Regen Res 12(10):1610-1612.

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Open peer reviewer: Jigar Pravinchandra Modi, Florida Atlantic University, USA.

Bélanger M, Allaman I, Magistretti PJ (2011) Brain energy metabolism: focus on astrocyte-neuron metabolic cooperation. Cell Metab 14:724-738.

Cabilly Y, Barbi M, Geva M, Marom L, Chetrit D, Ehrlich M,Elroy-Stein O (2012) Poor cerebral inflammatory response in eIF2B knock-in mice: implications for the aetiology of vanishing white matter disease. PLoS One 7:e46715.

Elroy-Stein O, Schiffmann R (2015) Vanishing White Matter Disease. In: Rosenberg’s Molecular and Genetic Basis of Neurological and Psychiatric Disease, 5thed (Rosenberg RN, Pascual JM,eds), pp 1015-1030. London, UK: Elsevier Inc.

Gat-Viks I, Geiger T, Barbi M, Raini G, Elroy-Stein O (2015) Proteomics-level analysis of myelin formation and regeneration in a mouse model for Vanishing White Matter disease. J Neurochem 134:513-526.

Geva M, Cabilly Y, Assaf Y, Mindroul N, Marom L, Raini G, Pinchasi D, Elroy-Stein O (2010) A mouse model for eukaryotic translation initiation factor 2B-leucodystrophy reveals abnormal development of brain white matter. Brain 133:2448-2461.

Kantor L, Harding HP, Ron D, Schiffmann R, Kaneski CR, Kimball SR, Elroy-Stein O (2005) Heightened stress response in primary fibroblasts expressing mutant eIF2B genes from CACH/VWM leukodystrophy patients. Hum Genet 118:99-106.

Leegwater PA, Vermeulen G, Könst 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.

Raini G, Sharet R, Herrero M, Atzmon A, Shenoy A, Geiger T,Elroy-Stein O (2017) Mutant eIF2B leads to impaired mitochondrial oxidative phosphorylation in vanishing white matter disease. J Neurochem 141:694-707.

Schoenfeld R, Wong A, Silva J, Li M, Itoh A, Horiuchi M, Itoh T,Pleasure D, Cortopassi G (2010) Oligodendroglial differentiation induces mitochondrial genes and inhibition of mitochondrial function represses oligodendroglial differentiation. Mitochondrion 10:143-150.

Wek RC, Jiang HY, Anthony TG (2006) Coping with stress: eIF2 kinases and translational control. Biochem Soc Trans 34:7-11.

Young SK, Wek RC (2016) Upstream open reading frames differentially regulate gene-specific translation in the integrated stress response. J Biol Chem 291:16927-16935.

10.4103/1673-5374.217329