Glyceraldehyde-3-Phosphate Dehydrogenase Facilitates Macroautophagic Degradation of Mutant Huntingtin Protein Aggregates.
Autophagy
/ physiology
Cell Line, Tumor
Gene Knockdown Techniques
Glyceraldehyde-3-Phosphate Dehydrogenases
/ antagonists & inhibitors
HEK293 Cells
Humans
Huntingtin Protein
/ genetics
Neuroblastoma
Peptides
/ genetics
Protein Aggregates
RNA Interference
RNA, Small Interfering
/ genetics
Ras Homolog Enriched in Brain Protein
/ metabolism
TOR Serine-Threonine Kinases
/ metabolism
Autophagy
Clearance
Multifunctional protein
Neurodegenerative disorder
mTOR pathway
Journal
Molecular neurobiology
ISSN: 1559-1182
Titre abrégé: Mol Neurobiol
Pays: United States
ID NLM: 8900963
Informations de publication
Date de publication:
Nov 2021
Nov 2021
Historique:
received:
01
04
2021
accepted:
11
08
2021
pubmed:
19
8
2021
medline:
25
3
2022
entrez:
18
8
2021
Statut:
ppublish
Résumé
Protein aggregate accumulation is a pathological hallmark of several neurodegenerative disorders. Autophagy is critical for clearance of aggregate-prone proteins. In this study, we identify a novel role of the multifunctional glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in clearance of intracellular protein aggregates. Previously, it has been reported that though clearance of wild-type huntingtin protein is mediated by chaperone-mediated autophagy (CMA), however, degradation of mutant huntingtin (mHtt with numerous poly Q repeats) remains impaired by this route as mutant Htt binds with high affinity to Hsc70 and LAMP-2A. This delays delivery of misfolded protein to lysosomes and results in accumulation of intracellular aggregates which are degraded only by macroautophagy. Earlier investigations also suggest that mHtt causes inactivation of mTOR signaling, causing upregulation of autophagy. GAPDH had earlier been reported to interact with mHtt resulting in cellular toxicity. Utilizing a cell culture model of mHtt aggregates coupled with modulation of GAPDH expression, we analyzed the formation of intracellular aggregates and correlated this with autophagy induction. We observed that GAPDH knockdown cells transfected with N-terminal mutant huntingtin (103 poly Q residues) aggregate-prone protein exhibit diminished autophagy. GAPDH was found to regulate autophagy via the mTOR pathway. Significantly more and larger-sized huntingtin protein aggregates were observed in GAPDH knockdown cells compared to empty vector-transfected control cells. This correlated with the observed decrease in autophagy. Overexpression of GAPDH had a protective effect on cells resulting in a decreased load of aggregates. Our results demonstrate that GAPDH assists in the clearance of protein aggregates by autophagy induction. These findings provide a new insight in understanding the mechanism of mutant huntingtin aggregate clearance. By studying the molecular mechanism of protein aggregate clearance via GAPDH, we hope to provide a new approach in targeting and understanding several neurodegenerative disorders.
Identifiants
pubmed: 34406601
doi: 10.1007/s12035-021-02532-5
pii: 10.1007/s12035-021-02532-5
doi:
Substances chimiques
HTT protein, human
0
Huntingtin Protein
0
Peptides
0
Protein Aggregates
0
RHEB protein, human
0
RNA, Small Interfering
0
Ras Homolog Enriched in Brain Protein
0
polyglutamine
26700-71-0
Glyceraldehyde-3-Phosphate Dehydrogenases
EC 1.2.1.-
MTOR protein, human
EC 2.7.1.1
TOR Serine-Threonine Kinases
EC 2.7.11.1
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
5790-5798Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
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