title: GGGGCC repeat expansion in C9orf72 compromises nucleocytoplasmic transport.
abstract: The GGGGCC (G4C2) repeat expansion in a noncoding region of C9orf72 is the most common cause of sporadic and familial forms of amyotrophic lateral sclerosis and frontotemporal dementia. The basis for pathogenesis is unknown. To elucidate the consequences of G4C2 repeat expansion in a tractable genetic system, we generated transgenic fly lines expressing 8, 28 or 58 G4C2-repeat-containing transcripts that do not have a translation start site (AUG) but contain an open-reading frame for green fluorescent protein to detect repeat-associated non-AUG (RAN) translation. We show that these transgenic animals display dosage-dependent, repeat-length-dependent degeneration in neuronal tissues and RAN translation of dipeptide repeat (DPR) proteins, as observed in patients with C9orf72-related disease. This model was used in a large-scale, unbiased genetic screen, ultimately leading to the identification of 18 genetic modifiers that encode components of the nuclear pore complex (NPC), as well as the machinery that coordinates the export of nuclear RNA and the import of nuclear proteins. Consistent with these results, we found morphological abnormalities in the architecture of the nuclear envelope in cells expressing expanded G4C2 repeats in vitro and in vivo. Moreover, we identified a substantial defect in RNA export resulting in retention of RNA in the nuclei of Drosophila cells expressing expanded G4C2 repeats and also in mammalian cells, including aged induced pluripotent stem-cell-derived neurons from patients with C9orf72-related disease. These studies show that a primary consequence of G4C2 repeat expansion is the compromise of nucleocytoplasmic transport through the nuclear pore, revealing a novel mechanism of neurodegeneration.
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Brian D
authors: BD
authors: Freibaum
authors: Department of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Yubing
authors: Y
authors: Lu
authors: Department of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Rodrigo
authors: R
authors: Lopez-Gonzalez
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Nam Chul
authors: NC
authors: Kim
authors: Department of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Sandra
authors: S
authors: Almeida
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Kyung-Ha
authors: KH
authors: Lee
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Nisha
authors: N
authors: Badders
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Marc
authors: M
authors: Valentine
authors: Memory and Aging Center, Department of Neurology, University of California San Francisco, San Francisco, California 94158, USA.
authors: Bruce L
authors: BL
authors: Miller
authors: Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors: Philip C
authors: PC
authors: Wong
authors: Department of Neuroscience, Mayo Clinic Florida, Jacksonville, Florida 32224, USA.
authors: Leonard
authors: L
authors: Petrucelli
authors: Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: Hong Joo
authors: HJ
authors: Kim
authors: Department of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors: Fen-Biao
authors: FB
authors: Gao
authors: Howard Hughes Medical Institute, Department of Cell and Molecular Biology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors: J Paul
authors: JP
authors: Taylor
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000235:genetics*; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000473:pathology; D000818:Animals; D030801:Animals, Genetically Modified; D000073885:C9orf72 Protein; D042622:DNA Repeat Expansion / Q000235:genetics*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D005123:Eye / Q000378:metabolism; D005260:Female; D057180:Frontotemporal Dementia / Q000235:genetics / Q000473:pathology; D006367:HeLa Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D008297:Male; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D022022:Nuclear Pore / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016366:Open Reading Frames / Q000235:genetics*; D010641:Phenotype; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D034443:RNA Transport / Q000235:genetics*; D012469:Salivary Glands / Q000166:cytology / Q000378:metabolism / Q000473:pathology
chemical_list: D000073885:C9orf72 Protein; C568651:C9orf72 protein, human; D011506:Proteins; D012313:RNA