@article{f718c637226546af8fecae70fc82e814,
title = "Mutant FUS and ELAVL4 (HuD) Aberrant Crosstalk in Amyotrophic Lateral Sclerosis",
abstract = "Amyotrophic lateral sclerosis (ALS) has been genetically linked to mutations in RNA-binding proteins (RBPs), including FUS. Here, we report the RNA interactome of wild-type and mutant FUS in human motor neurons (MNs). This analysis identified a number of RNA targets. Whereas the wild-type protein preferentially binds introns, the ALS mutation causes a shift toward 3' UTRs. Neural ELAV-like RBPs are among mutant FUS targets. As a result, ELAVL4 protein levels are increased in mutant MNs. ELAVL4 and mutant FUS interact and co-localize in cytoplasmic speckles with altered biomechanical properties. Upon oxidative stress, ELAVL4 and mutant FUS are engaged in stress granules. In the spinal cord of FUS ALS patients, ELAVL4 represents a neural-specific component of FUS-positive cytoplasmic aggregates, whereas in sporadic patients it co-localizes with phosphorylated TDP-43-positive inclusions. We propose that pathological mutations in FUS trigger an aberrant crosstalk with ELAVL4 with implications for ALS.",
keywords = "amytrophic lateral sclerosis, Brillouin, ELAVL4, FUS, HuD, motor neuron, PAR-CLIP, RNA-binding protein, stress granules, TDP-43",
author = "\{De Santis\}, Riccardo and Vincenzo Alfano and \{de Turris\}, Valeria and Alessio Colantoni and Laura Santini and Garone, \{Maria Giovanna\} and Giuseppe Antonacci and Giovanna Peruzzi and Emma Sudria-Lopez and Emanuel Wyler and Anink, \{Jasper J.\} and Eleonora Aronica and Markus Landthaler and Pasterkamp, \{R. Jeroen\} and Irene Bozzoni and Alessandro Rosa",
note = "Funding Information: The authors wish to thank the Imaging Facility and the Flow Cytometry Facility at Center for Life Nano Science, Istituto Italiano di Tecnologia, for support and technical advice. We thank Dr. Giulia M.R. De Luca (Scientific Volume Imaging, BV) for helpful discussion on deconvolution with the Huygens software. This work was partially supported by AriSLA pilot grant 2016 “ StressFUS ” to A.R.; ERC-2013 ( AdG 340172–MUNCODD ), AriSLA full grant 2014 “ ARCI ,” Human Frontiers Science Program Award RGP0009/2014 , and Epigen-Epigenomics Flagship Project and Telethon ( GGP16213 ) to I.B.; ALS Stichting grants TOTALS and FUNCTIONALS, Prinses Beatrix Spierfonds W.OR15-17 to R.J.P.; and ALS Stichting grant “The Dutch ALS Tissue Bank” to E.A. We acknowledge the team who helped in the collection of ALS tissue samples (Prof. Dr. D. Troost, Prof. Dr. M. de Visser, Dr. A.J. van der Kooi, and Dr. J. Raaphorst). Funding Information: The authors wish to thank the Imaging Facility and the Flow Cytometry Facility at Center for Life Nano Science, Istituto Italiano di Tecnologia, for support and technical advice. We thank Dr. Giulia M.R. De Luca (Scientific Volume Imaging, BV) for helpful discussion on deconvolution with the Huygens software. This work was partially supported by AriSLA pilot grant 2016 “StressFUS” to A.R.; ERC-2013 (AdG 340172–MUNCODD), AriSLA full grant 2014 “ARCI,” Human Frontiers Science Program Award RGP0009/2014, and Epigen-Epigenomics Flagship Project and Telethon (GGP16213) to I.B.; ALS Stichting grants TOTALS and FUNCTIONALS, Prinses Beatrix Spierfonds W.OR15-17 to R.J.P.; and ALS Stichting grant “The Dutch ALS Tissue Bank” to E.A. We acknowledge the team who helped in the collection of ALS tissue samples (Prof. Dr. D. Troost, Prof. Dr. M. de Visser, Dr. A.J. van der Kooi, and Dr. J. Raaphorst). A.R. R.D.S. and V.A. conceived the project, designed the work, and analyzed the results. R.D.S. performed the PAR-CLIP, the reporter assays and the coIP, and contributed to the bioinformatics analysis and paper writing. V.A. cultured and differentiated MNs, quantified ELAVL4 levels in MNs, performed the stainings, and contributed to paper writing. V.d.T. acquired and analyzed microscopy images and performed quantitative analysis of ELAVL4 from confocal images. A.C. performed the bioinformatics analysis of the PAR-CLIP. L.S. contributed to cell culture and MN differentiation and to the PAR-CLIP. M.G.G. contributed to the generation of plasmids, cell culture, and MN differentiation. G.A. performed BDB acquisition and analysis. G.P. performed MNs sorting. J.J.A. and E.A. provided patients{\textquoteright} tissues and immunohistochemistry on sporadic and FUS ALS patients{\textquoteright} samples. E.S.-L. and R.J.P. contributed to the staining on FUS patients{\textquoteright} samples. E.W. contributed to PAR-CLIP and bioinformatics analysis. M.L. coordinated the PAR-CLIP work. I.B. provided critical feedback and helped shape the research. A.R. coordinated the work and wrote the paper. The authors declare no competing interests. Funding Information: The Genotype-Tissue Expression (GTEx) Project was supported by the Common Fund of the Office of the Director of the National Institutes of Health, and by NCI, NHGRI, NHLBI, NIDA, NIMH, and NINDS. The data used for the analyses described in this manuscript were obtained from the GTEx Portal ( https://www.gtexportal.org/home/ ), Version 7, on 07/31/18. Publisher Copyright: {\textcopyright} 2019 The Author(s)",
year = "2019",
month = jun,
day = "25",
doi = "10.1016/j.celrep.2019.05.085",
language = "English",
volume = "27",
pages = "3818--3831.e5",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Elsevier",
number = "13",
}