ALS/FTD mutations in UBQLN2 impede autophagy by reducing autophagosome acidification through loss of function

Josephine J. Wu, Ashley Cai, Jessie E. Greenslade, Nicole R. Higgins, Cong Fan, Nhat T.T. Le, Micaela Tatman, Alexandra M. Whiteley, Miguel A. Prado, Birger V. Dieriks, Maurice A. Curtis, Christopher E. Shaw, Teepu Siddique, Richard L.M. Faull, Emma L. Scotter, Daniel Finley, Mervyn J. Monteiro*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

49 Scopus citations


Mutations in UBQLN2 cause amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and other neurodegenerations. However, the mechanism by which the UBQLN2 mutations cause disease remains unclear. Alterations in proteins involved in autophagy are prominent in neuronal tissue of human ALS UBQLN2 patients and in a transgenic P497S UBQLN2 mouse model of ALS/FTD, suggesting a pathogenic link. Here, we show UBQLN2 functions in autophagy and that ALS/FTD mutant proteins compromise this function. Inactivation of UBQLN2 expression in HeLa cells reduced autophagic flux and autophagosome acidification. The defect in acidification was rescued by reexpression of wild type (WT) UBQLN2 but not by any of the five different UBQLN2 ALS/FTD mutants tested. Proteomic analysis and immunoblot studies revealed P497S mutant mice and UBQLN2 knockout HeLa and NSC34 cells have reduced expression of ATP6v1g1, a critical subunit of the vacuolar ATPase (V-ATPase) pump. Knockout of UBQLN2 expression in HeLa cells decreased turnover of ATP6v1g1, while overexpression of WT UBQLN2 increased biogenesis of ATP6v1g1 compared with P497S mutant UBQLN2 protein. In vitro interaction studies showed that ATP6v1g1 binds more strongly to WT UBQLN2 than to ALS/FTD mutant UBQLN2 proteins. Intriguingly, overexpression of ATP6v1g1 in UBQLN2 knockout HeLa cells increased autophagosome acidification, suggesting a therapeutic approach to overcome the acidification defect. Taken together, our findings suggest that UBQLN2 mutations drive pathogenesis through a dominant-negative loss-of-function mechanism in autophagy and that UBQLN2 functions as an important regulator of the expression and stability of ATP6v1g1. These findings may have important implications for devising therapies to treat UBQLN2-linked ALS/FTD.

Original languageEnglish (US)
Pages (from-to)15230-15241
Number of pages12
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number26
StatePublished - Jun 30 2020


  • Autophagy
  • UBQLN2
  • Ubiquilin
  • Vacuolar ATPase pump
  • amyotrophic lateral sclerosis

ASJC Scopus subject areas

  • General


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