ALS-associated mutations in FUS disrupt the axonal distribution and function of SMN

• Published in: Human molecular genetics Vol. 22; no. 18; pp. 3690 - 3704
• Main Authors: Groen, Ewout J N, Fumoto, Katsumi, Blokhuis, Anna M, Engelen-Lee, Jooyeon, Zhou, Yeping, van den Heuvel, Dianne M A, Koppers, Max, van Diggelen, Femke, van Heest, Jessica, Demmers, Jeroen A A, Kirby, Janine, Shaw, Pamela J, Aronica, Eleonora, Spliet, Wim G M, Veldink, Jan H, van den Berg, Leonard H, Pasterkamp, R Jeroen
• Format: Journal Article
• Language: English
• Published: England 15.09.2013
• Subjects: Amyotrophic Lateral Sclerosis - genetics; Amyotrophic Lateral Sclerosis - metabolism; Animals; Axons - metabolism; Axons - ultrastructure; Cell Line, Tumor; Gene Expression; Growth Cones - ultrastructure; Humans; Mice; Mice, Inbred C57BL; Motor Neurons - metabolism; Motor Neurons - ultrastructure; Mutation; Phenotype; RNA-Binding Protein FUS - chemistry; RNA-Binding Protein FUS - genetics; RNA-Binding Protein FUS - metabolism; Survival of Motor Neuron 1 Protein - chemistry; Survival of Motor Neuron 1 Protein - genetics; Survival of Motor Neuron 1 Protein - metabolism; Transfection
• ISSN: 0964-6906; 1460-2083
• DOI: 10.1093/hmg/ddt222

Mutations in the RNA binding protein fused in sarcoma/translated in liposarcoma (FUS/TLS) cause amyotrophic lateral sclerosis (ALS). Although ALS-linked mutations in FUS often lead to a cytosolic mislocalization of the protein, the pathogenic mechanisms underlying these mutations remain poorly understood. To gain insight into these mechanisms, we examined the biochemical, cell biological and functional properties of mutant FUS in neurons. Expression of different FUS mutants (R521C, R521H, P525L) in neurons caused axonal defects. A protein interaction screen performed to explain these phenotypes identified numerous FUS interactors including the spinal muscular atrophy (SMA) causing protein survival motor neuron (SMN). Biochemical experiments showed that FUS and SMN interact directly and endogenously, and that this interaction can be regulated by FUS mutations. Immunostaining revealed co-localization of mutant FUS aggregates and SMN in primary neurons. This redistribution of SMN to cytosolic FUS accumulations led to a decrease in axonal SMN. Finally, cell biological experiments showed that overexpression of SMN rescued the axonal defects induced by mutant FUS, suggesting that FUS mutations cause axonal defects through SMN. This study shows that neuronal aggregates formed by mutant FUS protein may aberrantly sequester SMN and concomitantly cause a reduction of SMN levels in the axon, leading to axonal defects. These data provide a functional link between ALS-linked FUS mutations, SMN and neuronal connectivity and support the idea that different motor neuron disorders such as SMA and ALS may be caused, in part, by defects in shared molecular pathways.