TY - JOUR
T1 - C9orf72 arginine-rich dipeptide repeat proteins disrupt karyopherin-mediated nuclear import
AU - Hayes, Lindsey R.
AU - Duan, Lauren
AU - Bowen, Kelly
AU - Kalab, Petr
AU - Rothstein, Jeffrey D.
N1 - Funding Information:
National Institute of Neurological Disorders and Stroke K08NS104273 Lindsey R. Hayes National Institute of Neurological Disorders and Stroke R01NS094239 Jeffrey D Rothstein National Institute of Neurological Disorders and Stroke P01NS099114 Jeffrey D Rothstein National Institute on Aging RF1AG062171 Jeffrey D Rothstein The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© Hayes et al.
PY - 2020/3
Y1 - 2020/3
N2 - Disruption of nucleocytoplasmic transport is increasingly implicated in the pathogenesis of neurodegenerative diseases, including ALS caused by a C9orf72 hexanucleotide repeat expansion. However, the mechanism(s) remain unclear. Karyopherins, including importin b and its cargo adaptors, have been shown to co-precipitate with the C9orf72 arginine-containing dipeptide repeat proteins (R-DPRs), poly-glycine arginine (GR) and poly-proline arginine (PR), and are protective in genetic modifier screens. Here, we show that R-DPRs interact with importin b, disrupt its cargo loading, and inhibit nuclear import of importin b, importin a/b, and transportin cargoes in permeabilized mouse neurons and HeLa cells, in a manner that can be rescued by RNA. Although R-DPRs induce widespread protein aggregation in this in vitro system, transport disruption is not due to nucleocytoplasmic transport protein sequestration, nor blockade of the phenylalanine-glycine (FG)-rich nuclear pore complex. Our results support a model in which R-DPRs interfere with cargo loading on karyopherins.
AB - Disruption of nucleocytoplasmic transport is increasingly implicated in the pathogenesis of neurodegenerative diseases, including ALS caused by a C9orf72 hexanucleotide repeat expansion. However, the mechanism(s) remain unclear. Karyopherins, including importin b and its cargo adaptors, have been shown to co-precipitate with the C9orf72 arginine-containing dipeptide repeat proteins (R-DPRs), poly-glycine arginine (GR) and poly-proline arginine (PR), and are protective in genetic modifier screens. Here, we show that R-DPRs interact with importin b, disrupt its cargo loading, and inhibit nuclear import of importin b, importin a/b, and transportin cargoes in permeabilized mouse neurons and HeLa cells, in a manner that can be rescued by RNA. Although R-DPRs induce widespread protein aggregation in this in vitro system, transport disruption is not due to nucleocytoplasmic transport protein sequestration, nor blockade of the phenylalanine-glycine (FG)-rich nuclear pore complex. Our results support a model in which R-DPRs interfere with cargo loading on karyopherins.
UR - http://www.scopus.com/inward/record.url?scp=85080991253&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85080991253&partnerID=8YFLogxK
U2 - 10.7554/eLife.51685
DO - 10.7554/eLife.51685
M3 - Article
C2 - 32119645
AN - SCOPUS:85080991253
VL - 9
JO - eLife
JF - eLife
SN - 2050-084X
M1 - e51685
ER -