RT Journal Article
SR Electronic
T1 O-GlcNAcylation regulates neurofilament-light assembly and function and is perturbed by Charcot-Marie-Tooth disease mutations
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2023.02.22.529563
DO 10.1101/2023.02.22.529563
A1 Duc T. Huynh
A1 Jimin Hu
A1 Jordan R. Schneider
A1 Kalina N. Tsolova
A1 Erik J. Soderblom
A1 Abigail J. Watson
A1 Jen-Tsan Chi
A1 Chantell S. Evans
A1 Michael Boyce
YR 2023
UL http://biorxiv.org/content/early/2023/02/22/2023.02.22.529563.abstract
AB The neurofilament (NF) cytoskeleton is critical for neuronal morphology and function. In particular, the neurofilament-light (NF-L) subunit is required for NF assembly in vivo and is mutated in subtypes of Charcot-Marie-Tooth (CMT) disease. NFs are highly dynamic, and the regulation of NF assembly state is incompletely understood. Here, we demonstrate that human NF-L is modified in a nutrient-sensitive manner by O-linked-β-N-acetylglucosamine (O-GlcNAc), a ubiquitous form of intracellular glycosylation. We identify five NF-L O-GlcNAc sites and show that they regulate NF assembly state. Interestingly, NF-L engages in O-GlcNAc-mediated protein-protein interactions with itself and with the NF component α-internexin, implying that O-GlcNAc is a general regulator of NF architecture. We further show that NF-L O-GlcNAcylation is required for normal organelle trafficking in primary neurons, underlining its functional significance. Finally, several CMT-causative NF-L mutants exhibit perturbed O-GlcNAc levels and resist the effects of O-GlcNAcylation on NF assembly state, indicating a potential link between dysregulated O-GlcNAcylation and pathological NF aggregation. Our results demonstrate that site-specific glycosylation regulates NF-L assembly and function, and aberrant NF O-GlcNAcylation may contribute to CMT and other neurodegenerative disorders.Competing Interest StatementThe authors have declared no competing interest.