FUS is Phosphorylated by DNA-PK and Accumulates in the Cytoplasm after DNA Damage

• Type of Material: Article
• Authors: Qiudong Deng, Emory University; Christopher J. Holler, Emory University; Georgia Taylor, Emory University; Kathryn F. Hudson, Emory University; William Watkins, Emory University; Marla Gearing, Emory University; Daisuke Ito, Keio University; Melissa E. Murray, Mayo Clinic; Dennis W. Dickson, Mayo Clinic; Nicholas Seyfried, Emory University; Thomas Kukar, Emory University
• Language: English
• Date: 2014-06-04
• Publisher: Society for Neuroscience
• Publication Version: Final Published Version
• Copyright Statement: © 2014 the authors
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1523/JNEUROSCI.0172-14.2014
• Title of Journal or Parent Work: Journal of Neuroscience Nursing
• ISSN: 0270-6474
• Volume: 34
• Issue: 23
• Start Page: 7802
• End Page: 7813
• Grant/Funding Information: This work was supported by the National Institutes of Health (Grants P30NS069289, P50AG032362, and R00AG032362 to T.K. and Training Grant T32 “Training and translational research in Neurology” 2T32 NS 007480 to Q.D. and C.J.H.) and the Alzheimer's Association (New Investigator Research Grant to T.K.).
• Keywords: Fused in Sarcoma (FUS); B-LYMPHOCYTE DEVELOPMENT; Neurosciences & Neurology; Life Sciences & Biomedicine; ANALYSIS REVEALS; CELL-LINE; Neurosciences; frontotemporal lobar degeneration (FTLD); NUCLEAR IMPORT; EWING SARCOMA; FET PROTEINS; FTLD-FUS; cytoplasmic translocation; Science & Technology; AMYOTROPHIC-LATERAL-SCLEROSIS; DEPENDENT PROTEIN-KINASE; phosphorylation; DNA damage; FRONTOTEMPORAL LOBAR DEGENERATION; amyotrophic lateral sclerosis (ALS)
• Research Categories: Health Sciences, Pathology; Health Sciences, Pharmacology

Abstract

Abnormal cytoplasmic accumulation of Fused in Sarcoma (FUS) in neurons defines subtypes of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). FUS is a member of the FET protein family that includes Ewing's sarcoma (EWS) and TATA-binding protein-associated factor 2N (TAF15). FET proteins are predominantly localized to the nucleus, where they bind RNA and DNA to modulate transcription, mRNA splicing, and DNA repair. In ALS cases with FUS inclusions (ALS-FUS), mutations in the FUS gene cause disease, whereas FTLD cases with FUS inclusions (FTLD-FUS) do not harbor FUS mutations. Notably, in FTLD-FUS, all FET proteins accumulate with their nuclear import receptor Transportin 1 (TRN1), in contrast ALS-FUS inclusions are exclusively positive for FUS. In the present study, we show that induction of DNA damage replicates several pathologic hallmarks of FTLD-FUS in immortalized human cells and primary human neurons and astrocytes. Treatment with the antibiotic calicheamicin γ1, which causes DNA double-strand breaks, leads to the cytoplasmic accumulation of FUS, TAF15, EWS, and TRN1. Moreover, cytoplasmic translocation of FUS is mediated by phosphorylation of its N terminus by the DNA-dependent protein kinase. Finally, we observed elevated levels of phospho-H2AX in FTLD-FUS brains, indicating that DNA damage occurs in patients. Together, our data reveal a novel regulatory mechanism for FUS localization in cells and suggest that DNA damage may contribute to the accumulation of FET proteins observed in human FTLD-FUS cases, but not in ALS-FUS. © 2014 the authors.