FUS regulates RAN translation through modulating the G-quadruplex structure of GGGGCC repeat RNA in C9orf72-linked ALS/FTD

Yuzo Fujino; Morio Ueyama; Taro Ishiguro; Daisaku Ozawa; Hayato Ito; Toshihiko Sugiki; Asako Murata; Akira Ishiguro; Tania Gendron; Kohji Mori; Eiichi Tokuda; Tomoya Taminato; Takuya Konno; Akihide Koyama; Yuya Kawabe; Toshihide Takeuchi; Yoshiaki Furukawa; Toshimichi Fujiwara; Manabu Ikeda; Toshiki Mizuno; Hideki Mochizuki; Hidehiro Mizusawa; Keiji Wada; Kinya Ishikawa; Osamu Onodera; Kazuhiko Nakatani; Leonard Petrucelli; Hideki Taguchi; Yoshitaka Nagai

doi:10.7554/eLife.84338

FUS regulates RAN translation through modulating the G-quadruplex structure of GGGGCC repeat RNA in C9orf72-linked ALS/FTD

Elife. 2023 Jul 18:12:RP84338. doi: 10.7554/eLife.84338.

Authors

Yuzo Fujino^{1

2}, Morio Ueyama^{1

3

4}, Taro Ishiguro^{4

5}, Daisaku Ozawa^{1

3}, Hayato Ito⁶, Toshihiko Sugiki⁷, Asako Murata⁸, Akira Ishiguro⁹, Tania Gendron¹⁰, Kohji Mori¹¹, Eiichi Tokuda¹², Tomoya Taminato^{1

3}, Takuya Konno¹³, Akihide Koyama¹³, Yuya Kawabe¹¹, Toshihide Takeuchi^{3

14}, Yoshiaki Furukawa¹², Toshimichi Fujiwara⁷, Manabu Ikeda¹¹, Toshiki Mizuno², Hideki Mochizuki¹⁵, Hidehiro Mizusawa⁵, Keiji Wada⁴, Kinya Ishikawa⁵, Osamu Onodera¹³, Kazuhiko Nakatani⁸, Leonard Petrucelli¹⁰, Hideki Taguchi^{6

16}, Yoshitaka Nagai^{1

3

4

15}

Affiliations

¹ Department of Neurology, Kindai University Faculty of Medicine, Osaka-Sayama, Japan.
² Department of Neurology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
³ Department of Neurotherapeutics, Osaka University Graduate School of Medicine, Suita, Japan.
⁴ Department of Degenerative Neurological Diseases, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan.
⁵ Department of Neurology and Neurological Science, Tokyo Medical and Dental University, Tokyo, Japan.
⁶ School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
⁷ Laboratory of Molecular Biophysics, Institute for Protein Research, Osaka University, Osaka, Japan.
⁸ Department of Regulatory Bioorganic Chemistry, The Institute of Scientific and28 Industrial Research, Osaka University, Osaka, Japan.
⁹ Research Center for Micro-nano Technology, Hosei University, Tokyo, Japan.
¹⁰ Department of Neuroscience, Mayo Clinic, Jacksonville, United States.
¹¹ Department of Psychiatry, Osaka University Graduate School of Medicine, Osaka, Japan.
¹² Department of Chemistry, Keio University, Kanagawa, Japan.
¹³ Department of Neurology, Clinical Neuroscience Branch, Brain Research Institute, Niigata University, Niigata, Japan.
¹⁴ Life Science Research Institute, Kindai University, Osaka, Japan.
¹⁵ Department of Neurology, Osaka University Graduate School of Medicine, Osaka, Japan.
¹⁶ Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Kanagawa, Japan.

Abstract

Abnormal expansions of GGGGCC repeat sequence in the noncoding region of the C9orf72 gene is the most common cause of familial amyotrophic lateral sclerosis and frontotemporal dementia (C9-ALS/FTD). The expanded repeat sequence is translated into dipeptide repeat proteins (DPRs) by noncanonical repeat-associated non-AUG (RAN) translation. Since DPRs play central roles in the pathogenesis of C9-ALS/FTD, we here investigate the regulatory mechanisms of RAN translation, focusing on the effects of RNA-binding proteins (RBPs) targeting GGGGCC repeat RNAs. Using C9-ALS/FTD model flies, we demonstrated that the ALS/FTD-linked RBP FUS suppresses RAN translation and neurodegeneration in an RNA-binding activity-dependent manner. Moreover, we found that FUS directly binds to and modulates the G-quadruplex structure of GGGGCC repeat RNA as an RNA chaperone, resulting in the suppression of RAN translation in vitro. These results reveal a previously unrecognized regulatory mechanism of RAN translation by G-quadruplex-targeting RBPs, providing therapeutic insights for C9-ALS/FTD and other repeat expansion diseases.

Keywords: ALS; C9orf72; D. melanogaster; FUS; RAN translation; RNA chaperone; neuroscience; repeat expansion disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amyotrophic Lateral Sclerosis* / pathology
C9orf72 Protein / genetics
C9orf72 Protein / metabolism
Drosophila / genetics
Frontotemporal Dementia* / pathology
Humans
RNA / metabolism
RNA-Binding Protein FUS / genetics
RNA-Binding Proteins / genetics

Substances

C9orf72 Protein
C9orf72 protein, human
FUS protein, human
RNA
RNA-Binding Protein FUS
RNA-Binding Proteins

Grants and funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.