Synergistic effect between proteasome and autophagosome in the clearance of polyubiquitinated TDP‐43

Urushitani, Makoto; Sato, Takashi; Bamba, Hitoshi; Hisa, Yasuo; Tooyama, Ikuo

doi:10.1002/jnr.22243

Cited by 114Amyotrophic Lateral Sclerosis ResearchAutophagy in Disease and TherapyUbiquitin and proteasome pathways

Synergistic effect between proteasome and autophagosome in the clearance of polyubiquitinated TDP‐43

Urushitani, Makoto, Sato, Takashi, Bamba, Hitoshi, Hisa, Yasuo, Tooyama, Ikuo·2010·Journal of Neuroscience Research

DOI 10.1002/jnr.22243

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Abstract

Cytoplasmic aggregates of ubiquitinated TAR DNA‐binding protein 43 (TDP‐43) are a pathological hallmark of amyotrophic lateral sclerosis (ALS). However, the mechanism of TDP‐43 polyubiquitination remains elusive. We investigated the effect of nuclear exclusion of TDP‐43 on aggregate formation and fragmentation, using TDP‐43 expression constructs for WT or mutant TDP‐43 with a modified nuclear localizing signal (LQ‐NLS). Overexpression of the LQ‐NLS mutant alone induced no detectable cytoplasmic aggregates during a 72‐hr period. Polyubiquitination of both WT TDP‐43 and the LQ‐NLS mutant was similar in total cell lysates exposed to the proteasome inhibitor lactacystin. However, analysis of subcellular fractions demonstrated a higher concentration of polyubiquitinated TDP‐43 in the nuclear fraction than in the cytosol for WT, and vice versa for the LQ‐NLS mutant. Polyubiquitin‐charged WT and mutant TDP‐43 were highly concentrated in the membrane/microsome fraction, which was also positive for the autophagosome marker LC3. In addition, the autophagy inhibitor 3‐methyladenine (3MA) blocked degradation of both TDP‐43 types, whereas lactacystin was minimally restorative. Furthermore, lactacystin plus 3MA induced prominent cytoplasmic aggregates. We also demonstrated mediation of TDP‐43 polyubiquitination by lysine 48 of ubiquitin, indicating a degradation signal in both TDP‐43 types. This is the first report delineating the distribution of polyubiquitinated TDP‐43 and the degradation pathway of TDP‐43 and clarifying the crucial role of autophagosomes in TDP‐43 clearance. We also demonstrate that nuclear exclusion itself is not an immediate trigger for ALS pathology. Further clarification of the mechanism of polyubiquitination of TDP‐43 and the role of autophagosomes may help in understanding and treating ALS. © 2009 Wiley‐Liss, Inc.

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“Inhibition of MTOR likewise reduces TARDBP aggregation and toxicity in multiple cell and animal models of ALS [353,358,359].”
From Autophagy and ALS: mechanistic insights and therapeutic implications (2022) · cites this paper as [358]

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BackgroundInfluential

Authors: Urushitani, M., Sato, T., Bamba, H., Hisa, Y., & Tooyama, I.
Affiliations: Shiga University of Medical Science (JP)
Kyoto Prefectural University of Medicine (JP)
Corresponding author: Urushitani, Makoto
ORCID: Urushitani, Makoto
Tooyama, Ikuo
Venue: Journal of Neuroscience Research
Publisher: Wiley
Published: 2009-10
Volume: 88
Issue: 4
Pages: 784-797
ISSN: 0360-4012 (ISSN-L)
1097-4547
DOI: 10.1002/jnr.22243
OpenAlex: W2104199101
Cited by: 114
References: 29
Citations by year: 2026: 7 · 2025: 4 · 2024: 1 · 2023: 7 · 2022: 6 · 2021: 5 · 2020: 8 · 2019: 7 · 2018: 7 · 2017: 7 · 2016: 8 · 2015: 3 · 2014: 10 · 2013: 5 · 2012: 9
FWCI: 6.17
Type: journal article
Language: en
Open access: closed
Licence: onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations: Journal of Neuroscience Research (publishedVersion)
PubMed (publishedVersion)
Topics: Health Sciences › Medicine › Neurology › Amyotrophic Lateral Sclerosis Research
Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
Keywords: TDP-43, polyubiquitination, amyotrophic lateral sclerosis, autophagy, proteasome, nuclear exclusion, cytoplasmic aggregates, protein degradation, LC3, 3-methyladenine, lactacystin
MeSH terms: Acetylcysteine, Adenine, Analysis of Variance, Autophagy, Cell Nucleus, DNA-Binding Proteins, Dose-Response Relationship, Drug, Drug Synergism, Humans, Mutation, Neuroblastoma, Subcellular Fractions, Transfection, Cysteine Proteinase Inhibitors, Protein Transport, Ubiquitin-Protein Ligases, Cell Line, Tumor, Proteasome Endopeptidase Complex, Green Fluorescent Proteins, Ubiquitination

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Synergistic effect between proteasome and autophagosome in the clearance of polyubiquitinated TDP‐43

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