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Arginine Methyltransferase PRMT8 Provides Cellular Stress Tolerance in Aging Motoneurons

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Arginine Methyltransferase PRMT8 Provides Cellular Stress Tolerance in Aging Motoneurons The Journal of Neuroscience, August 29, 2018 • 38(35):7683–7700 • 7683 Development/Plasticity/Repair Arginine Methyltransferase PRMT8 Provides Cellular Stress Tolerance in Aging Motoneurons X Zoltan Simandi,1,2 Krisztian Pajer,3 Katalin Karolyi,1 XTatiana Sieler,1 XLu-Lin Jiang,4 XZsuzsanna Kolostyak,2 X Zsanett Sari,2 Zoltan Fekecs,3 Attila Pap,2 XAndreas Patsalos,2 Gerardo Alvarado Contreras,5 XBalint Reho,6 Zoltan Papp,5 XXiufang Guo,7 XAttila Horvath,2 Greta Kiss,8 Zsolt Keresztessy,2 Gyo¨rgy Va´mosi,6 XJames Hickman,7 XHuaxi Xu,4 XDorothee Dormann,9,10 XTibor Hortobagyi,11 XMiklos Antal,8,12 Antal No´gra´di,3* and XLaszlo Nagy1,2,13* 1Sanford Burnham Prebys Medical Discovery Institute, Orlando, Florida 32827, 2Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary, HU 4032, 3Department of Anatomy, Histology and Embryology, University of Szeged, Szeged, Hungary, HU 6720, 4Neuroscience Initiative, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California 92037, 5Division of Clinical Physiology, Institute of Cardiology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary, HU 4032, 6Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary, HU 4032, 7NanoScience Technology Center, University of Central Florida, Orlando, Florida 32816, 8Department of Anatomy, Faculty of Medicine, University of Debrecen, Debrecen, Hungary, HU 4032, 9BioMedical Center, Ludwig-Maximilians-University Munich, Planegg-Martinsried, Germany 80539, 10Munich Cluster for Systems Neurology (SyNergy), Munich, Germany 80539, 11HAS-UD Cerebrovascular and Neurodegenerative Research Group, Department of Neurology and Neuropathology, University of Debrecen, Debrecen, Hungary, HU 4032, 12HAS-UD Neuroscience Research Group, University of Debrecen, Debrecen, Hungary, HU 4032, and 13HAS-UD Momentum Immunogenomics Research Group, University of Debrecen, Debrecen, Hungary, HU 4032 Aging contributes to cellular stress and neurodegeneration. Our understanding is limited regarding the tissue-restricted mechanisms providing protection in postmitotic cells throughout life. Here, we show that spinal cord motoneurons exhibit a high abundance of asymmetric dimethyl arginines (ADMAs) and the presence of this posttranslational modification provides protection against environ- mental stress. We identify protein arginine methyltransferase 8 (PRMT8) as a tissue-restricted enzyme responsible for proper ADMA levelinpostmitoticneurons.MalePRMT8knock-outmicedisplaydecreasedmusclestrengthwithagingduetoprematuredestabilization of neuromuscular junctions. Mechanistically, inhibition of methyltransferase activity or loss of PRMT8 results in accumulation of unrepaired DNA double-stranded breaks and decrease in the cAMP response-element-binding protein 1 (CREB1) level. As a conse- quence, the expression of CREB1-mediated prosurvival and regeneration-associated immediate early genes is dysregulated in aging PRMT8 knock-out mice. The uncovered role of PRMT8 represents a novel mechanism of stress tolerance in long-lived postmitotic neurons and identifies PRMT8 as a tissue-specific therapeutic target in the prevention of motoneuron degeneration. Key words: ADMA; aging; CREB1; motoneuron; neurodegeneration; PRMT8 Significance Statement Although most of the cells in our body have a very short lifespan, postmitotic neurons must survive for many decades. Longevity of a cell within the organism depends on its ability to properly regulate signaling pathways that counteract perturbations, such as DNA damage, oxidative stress, or protein misfolding. Here, we provide evidence that tissue-specific regulators of stress tolerance exist in postmitotic neurons. Specifically, we identify protein arginine methyltransferase 8 (PRMT8) as a cell-type-restricted arginine methyltransferase in spinal cord motoneurons (MNs). PRMT8-dependent arginine methylation is required for neuro- protection against age-related increased of cellular stress. Tissue-restricted expression and the enzymatic activity of PRMT8 make it an attractive target for drug development to delay the onset of neurodegenerative disorders. Introduction pathways (Haigis and Yankner, 2010). Accumulating evidence indi- The rate of cellular aging and the appearance of age-related pathol- cates that the longevity of a cell or organism depends on its ability to ogies are modulated by highly conserved stress response and repair properly regulate signaling pathways that counteract perturbations, such as protein misfolding, DNA damage, or oxidative stress (Kour- Received Nov. 29, 2017; revised June 22, 2018; accepted June 25, 2018. Author contributions: Z. Simandi and L.N. edited the paper; Z. Simandi, K.P., and L.N. designed research; research; Z.P., Z. Keresztessy, J.H., H.X., D.D., M.A., and A.N. contributed unpublished reagents/analytic tools; Z. Simandi, K.P., K.K., T.S., L.-L.J., Z. Kolostyak, Z. Sari, Z.F., A. Pap, A. Patsalos, G.A.C., B.R., X.G., and G.K. performed Z. Simandi, K.P., A.H., G.V., T.H., M.A., A.N., and L.N. analyzed data; Z. Simandi, A.N., and L.N. wrote the paper. 7684 • J. Neurosci., August 29, 2018 • 38(35):7683–7700 Simandi et al. • PRMT8 Provides Stress Tolerance in Neurons tis and Tavernarakis, 2011). However, stress responses must funda- PRMT8 is a unique member of the family because it shows highly mentally differ between proliferating cells, which can be easily tissue-specific expression by being restricted to the CNS (Taneda replaced, and cells that are terminally differentiated and nonreplace- et al., 2007; Kousaka et al., 2009). PRMT8 has been shown to act able, such as postmitotic neurons. In proliferating cells, the risk of as a posttranslational modifier of various proteins (Kim et al., propagating cellular defects is great, which can be potentially threat- 2008; Pahlich et al., 2008). Similar to PRMT1, its closest paralog, ening the survival of the entire organism. Therefore, elimination of PRMT8, is involved in the epigenetic control of gene expression damaged proliferating cells is the most straightforward solution. In and normal function of neurons (Simandi et al., 2015). However, contrast, in long-lived neurons, this could damage the integrity of the in vivo biological role of PRMT8 in the CNS and the mecha- the complex neural network, so alternative mechanisms need to be nisms resulting in neural defects remain largely unknown except used to properly maintain the homeostasis of these cells (Herrup and for very recent studies that described the role of PRMT8 in Pur- Yang, 2007; Kole et al., 2013). Identifying and understanding molec- kinje cells (Kim et al., 2015) and excitatory synaptic function ular mechanisms regulating cellular stress resistance in postmitotic (Penney et al., 2017). neurons is important for defining novel therapeutic targets in late- Here, we show that asymmetric dimethyl arginine (ADMA) onset neurodegenerative diseases. level declines during embryonic development in the mouse. Posttranslational modifications (PTMs), such as methylation, Strikingly, choline acetyltransferase (ChAT)ϩ MNs selectively acetylation, and phosphorylation are crucial for balanced func- maintain high ADMA level in the adult spinal cord. Fused in tions of stress response pathways and dysregulated PTMs can sarcoma (FUS), a prominent arginine methyltransferase target, potentially lead to a pathological state (Lake and Bedford, 2007; shows similar cell-type-restricted expression. Inhibition of meth- Dantuma and van Attikum, 2016). Arginine is a positively yltransferases results in accumulation of DNA double-stranded charged residue that is often found in protein motifs, including breaks (DSBs), altered FUS kinetics at DNA DSBs, and overall a the RGG/RG motifs (Thandapani et al., 2013). The RGG se- more vulnerable cellular state and decreased cellular viability. We quences within these motifs are substrate recognition sites for show that, among the asymmetric arginine methyltransferases, type I and type II protein arginine methyltransferases (PRMTs) PRMT8 is selectively expressed in the spinal cord MNs. Loss of catalyzing monomethylation and asymmetric or symmetric dim- PRMT8 results in a progressive decrease in muscle strength due to ethylation of arginine residues, respectively (Bedford and Clarke, the dysfunction and gradual loss of MNs in aging animals. The per- 2009; Blanc and Richard, 2017). In the last decade, hundreds of sistent stress in the absence of methyltransferase activity leads to methylarginine proteins have been detected by mass spectrome- decreased cAMP response-element-binding protein 1 (CREB1) level try and other proteomic and molecular techniques (Hart-Smith and insufficient activation of prosurvival and regeneration gene net- et al., 2012; Bremang et al., 2013; Guo et al., 2014; Auburger et al., work in response to aging-related oxidative and ER stress. This work 2016). These methyltransferase substrate proteins are involved in is a proof-of-concept showing that ADMA has a nonproliferation- transcriptional regulation (TAF15, CRTC2), pre-mRNA splicing associated, cell-type-restricted role and is required for proper stress (FUS, G3BP1, hnRNP A1, DDX3X, SAM68), mRNA translation response in postmitotic neurons. Based on our findings, PRMT8 (EIF3, EIF4), DNA-damage response (MRE11, 53BP1, FUS), and should be considered as a novel component of neural stress resis- regulation of apoptosis (PABP1). Prior in vitro studies estab- tance and a target for future drug discovery
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