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Giving protein folding a helping hand

The reversible phosphorylation of proteins controls virtually all aspects of cell and organismal function. Targeting phosphorylation offers a broad range of therapeutic opportunities, and thus kinases have become important therapeutic targets. As targets, phosphatases should be as attractive, but in fact they are more challenging to manipulate. Das et al. have found a safe and specific inhibitor, called Sephin1, that targets a regulatory subunit of protein phosphatase 1 in vivo. Sephin1 binds and inhibits PPP1R15A, but not the related regulatory phosphatase PPP1R15B. In mice, Sephin1 prolonged a stress-induced phospho-signaling pathway to prevent the pathological defects of the unrelated protein-misfolding diseases Charcot-Marie-Tooth 1B and amyotrophic lateral sclerosis.
Science, this issue p. 239

Abstract

Protein phosphorylation regulates virtually all biological processes. Although protein kinases are popular drug targets, targeting protein phosphatases remains a challenge. Here, we describe Sephin1 (selective inhibitor of a holophosphatase), a small molecule that safely and selectively inhibited a regulatory subunit of protein phosphatase 1 in vivo. Sephin1 selectively bound and inhibited the stress-induced PPP1R15A, but not the related and constitutive PPP1R15B, to prolong the benefit of an adaptive phospho-signaling pathway, protecting cells from otherwise lethal protein misfolding stress. In vivo, Sephin1 safely prevented the motor, morphological, and molecular defects of two otherwise unrelated protein-misfolding diseases in mice, Charcot-Marie-Tooth 1B, and amyotrophic lateral sclerosis. Thus, regulatory subunits of phosphatases are drug targets, a property exploited here to safely prevent two protein misfolding diseases.

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Supplementary Material

Summary

Materials and Methods
Figs. S1 to S6
References (2629)

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References and Notes

1
Harding H. P., Zhang Y., Bertolotti A., Zeng H., Ron D., Perk is essential for translational regulation and cell survival during the unfolded protein response. Mol. Cell 5, 897–904 (2000).
2
Scheuner D., Song B., McEwen E., Liu C., Laybutt R., Gillespie P., Saunders T., Bonner-Weir S., Kaufman R. J., Translational control is required for the unfolded protein response and in vivo glucose homeostasis. Mol. Cell 7, 1165–1176 (2001).
3
Harding H. P., Zhang Y., Zeng H., Novoa I., Lu P. D., Calfon M., Sadri N., Yun C., Popko B., Paules R., Stojdl D. F., Bell J. C., Hettmann T., Leiden J. M., Ron D., An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol. Cell 11, 619–633 (2003).
4
Tsaytler P., Harding H. P., Ron D., Bertolotti A., Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis. Science 332, 91–94 (2011).
5
Scheuner D., Patel R., Wang F., Lee K., Kumar K., Wu J., Nilsson A., Karin M., Kaufman R. J., Double-stranded RNA-dependent protein kinase phosphorylation of the alpha-subunit of eukaryotic translation initiation factor 2 mediates apoptosis. J. Biol. Chem. 281, 21458–21468 (2006).
6
Balch W. E., Morimoto R. I., Dillin A., Kelly J. W., Adapting proteostasis for disease intervention. Science 319, 916–919 (2008).
7
Holmes B., Brogden R. N., Heel R. C., Speight T. M., Avery G. S., Guanabenz. A review of its pharmacodynamic properties and therapeutic efficacy in hypertension. Drugs 26, 212–229 (1983).
8
Brostrom M. A., Lin X. J., Cade C., Gmitter D., Brostrom C. O., Loss of a calcium requirement for protein synthesis in pituitary cells following thermal or chemical stress. J. Biol. Chem. 264, 1638–1643 (1989).
9
Novoa I., Zhang Y., Zeng H., Jungreis R., Harding H. P., Ron D., Stress-induced gene expression requires programmed recovery from translational repression. EMBO J. 22, 1180–1187 (2003).
10
Harding H. P., Novoa I., Zhang Y., Zeng H., Wek R., Schapira M., Ron D., Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol. Cell 6, 1099–1108 (2000).
11
Meacham R. H., Ruelius H. W., Kick C. J., Peters J. R., Kocmund S. M., Sisenwine S. F., Wendt R. L., Relationship of guanabenz concentrations in brain and plasma to antihypertensive effect in the spontaneously hypertensive rat. J. Pharmacol. Exp. Ther. 214, 594–598 (1980).
12
Hall A. H., Smolinske S. C., Kulig K. W., Rumack B. H., Guanabenz overdose. Ann. Intern. Med. 102, 787–788 (1985).
13
Genoux D., Haditsch U., Knobloch M., Michalon A., Storm D., Mansuy I. M., Protein phosphatase 1 is a molecular constraint on learning and memory. Nature 418, 970–975 (2002).
14
Costa-Mattioli M., Gobert D., Stern E., Gamache K., Colina R., Cuello C., Sossin W., Kaufman R., Pelletier J., Rosenblum K., Krnjević K., Lacaille J. C., Nader K., Sonenberg N., eIF2α phosphorylation bidirectionally regulates the switch from short- to long-term synaptic plasticity and memory. Cell 129, 195–206 (2007).
15
Morris R. G. M., Garrud P., Rawlins J. N. P., O’Keefe J., Place navigation impaired in rats with hippocampal lesions. Nature 297, 681–683 (1982).
16
Pennuto M., Tinelli E., Malaguti M., Del Carro U., D’Antonio M., Ron D., Quattrini A., Feltri M. L., Wrabetz L., Ablation of the UPR-mediator CHOP restores motor function and reduces demyelination in Charcot-Marie-Tooth 1B mice. Neuron 57, 393–405 (2008).
17
D’Antonio M., Musner N., Scapin C., Ungaro D., Del Carro U., Ron D., Feltri M. L., Wrabetz L., Resetting translational homeostasis restores myelination in Charcot-Marie-Tooth disease type 1B mice. J. Exp. Med. 210, 821–838 (2013).
18
Wrabetz L., D’Antonio M., Pennuto M., Dati G., Tinelli E., Fratta P., Previtali S., Imperiale D., Zielasek J., Toyka K., Avila R. L., Kirschner D. A., Messing A., Feltri M. L., Quattrini A., Different intracellular pathomechanisms produce diverse Myelin Protein Zero neuropathies in transgenic mice. J. Neurosci. 26, 2358–2368 (2006).
19
Nordlund A., Oliveberg M., SOD1-associated ALS: A promising system for elucidating the origin of protein-misfolding disease. HFSP J. 2, 354–364 (2008).
20
Nishitoh H., Kadowaki H., Nagai A., Maruyama T., Yokota T., Fukutomi H., Noguchi T., Matsuzawa A., Takeda K., Ichijo H., ALS-linked mutant SOD1 induces ER stress- and ASK1-dependent motor neuron death by targeting Derlin-1. Genes Dev. 22, 1451–1464 (2008).
21
Wang L., Popko B., Roos R. P., An enhanced integrated stress response ameliorates mutant SOD1-induced ALS. Hum. Mol. Genet. 23, 2629–2638 (2014).
22
Gurney M. E., Pu H., Chiu A., Dal Canto M., Polchow C., Alexander D., Caliendo J., Hentati A., Kwon Y., Deng H., et, Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264, 1772–1775 (1994).
23
Marciniak S. J., Yun C. Y., Oyadomari S., Novoa I., Zhang Y., Jungreis R., Nagata K., Harding H. P., Ron D., CHOP induces death by promoting protein synthesis and oxidation in the stressed endoplasmic reticulum. Genes Dev. 18, 3066–3077 (2004).
24
Wang J., Farr G. W., Zeiss C. J., Rodriguez-Gil D. J., Wilson J. H., Furtak K., Rutkowski D. T., Kaufman R. J., Ruse C. I., Yates J. R., Perrin S., Feany M. B., Horwich A. L., Progressive aggregation despite chaperone associations of a mutant SOD1-YFP in transgenic mice that develop ALS. Proc. Natl. Acad. Sci. U.S.A. 106, 1392–1397 (2009).
25
Harding H. P., Zhang Y., Scheuner D., Chen J. J., Kaufman R. J., Ron D., Ppp1r15 gene knockout reveals an essential role for translation initiation factor 2 α (eIF2α) dephosphorylation in mammalian development. Proc. Natl. Acad. Sci. U.S.A. 106, 1832–1837 (2009).
26
Wang J., Farr G. W., Hall D. H., Li F., Furtak K., Dreier L., Horwich A. L., An ALS-linked mutant SOD1 produces a locomotor defect associated with aggregation and synaptic dysfunction when expressed in neurons of Caenorhabditis elegans. PLOS Genet. 5, e1000350 (2009).
27
Heiman-Patterson T. D., Deitch J. S., Blankenhorn E. P., Erwin K. L., Perreault M. J., Alexander B. K., Byers N., Toman I., Alexander G. M., Background and gender effects on survival in the TgN(SOD1-G93A)1Gur mouse model of ALS. J. Neurol. Sci. 236, 1–7 (2005).
28
Schaeffer V., Lavenir I., Ozcelik S., Tolnay M., Winkler D. T., Goedert M., Stimulation of autophagy reduces neurodegeneration in a mouse model of human tauopathy. Brain 135, 2169–2177 (2012).
29
Schindelin J., Arganda-Carreras I., Frise E., Kaynig V., Longair M., Pietzsch T., Preibisch S., Rueden C., Saalfeld S., Schmid B., Tinevez J. Y., White D. J., Hartenstein V., Eliceiri K., Tomancak P., Cardona A., Fiji: An open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).

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Published In

Science
Volume 348 | Issue 6231
10 April 2015

Submission history

Received: 8 December 2014
Accepted: 6 March 2015
Published in print: 10 April 2015

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Acknowledgments

We thank P. Tsaytler for some initial experiments on Sephin1; H. Meziane for the studies on memory; E. Fisher for SOD1G93A mice; E. Pettinato and C. Ferri for technical assistance; R. Roberts for discussions on CMT; A. Segonds-Pichon for statistical analysis; and members of the Bertolotti laboratory, M. Goedert, M. Hastings, and S. Munro for discussions. A.B. is an honorary fellow of the Clinical Neurosciences Department of Cambridge University. This work was supported by the Medical Research Council (UK) and the European Research Council (ERC) under the European Union’s Seventh Framework Programme (FP7/2007-2013)/ERC grant 309516. A.K. was supported by the European Molecular Biology Organization and Human Frontier Science Program, K.S. by the Swiss National Science Foundation, M.D. by the Italian Ministry of Health (GR-2011-02642791), and L.W. by NIH R01-NS55256. A.B. is a co-inventor on Great Britain patent WO 2014108520, covering benzylideneguanidine derivatives inhibitors of PPP1R15A. The data presented in this paper are tabulated in the main paper and the supplementary materials.

Authors

Affiliations

Indrajit Das
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
Agnieszka Krzyzosiak
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
Kim Schneider
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
Lawrence Wrabetz
Division of Genetics and Cell Biology, San Raffaele Scientific Institute, 20132 Milan, Italy.
Present address: Hunter James Kelly Research Institute (HJKRI), University at Buffalo School of Medicine and Biomedical Sciences, 701 Ellicott Street, Buffalo, NY 14203, USA.
Maurizio D’Antonio
Division of Genetics and Cell Biology, San Raffaele Scientific Institute, 20132 Milan, Italy.
Nicholas Barry
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
Anna Sigurdardottir
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
Anne Bertolotti [email protected]
Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.

Notes

Corresponding author. E-mail: [email protected]

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