Artificial tethering of LC3 or p62 to organelles is not sufficient to trigger autophagy.
Adaptor Proteins, Signal Transducing
/ genetics
Autophagy
/ drug effects
Biotin
/ metabolism
Cell Line, Tumor
Cytosol
/ metabolism
Endoplasmic Reticulum
/ metabolism
Golgi Apparatus
/ metabolism
Green Fluorescent Proteins
/ metabolism
Humans
Microtubule-Associated Proteins
/ genetics
Protein Binding
/ genetics
Protein Transport
/ genetics
RNA-Binding Proteins
/ genetics
Recombinant Proteins
/ chemistry
Streptavidin
/ metabolism
Journal
Cell death & disease
ISSN: 2041-4889
Titre abrégé: Cell Death Dis
Pays: England
ID NLM: 101524092
Informations de publication
Date de publication:
10 10 2019
10 10 2019
Historique:
received:
21
06
2019
accepted:
19
09
2019
revised:
12
09
2019
entrez:
12
10
2019
pubmed:
12
10
2019
medline:
28
8
2020
Statut:
epublish
Résumé
The retention using selective hooks (RUSH) system allows to retain a target protein fused to green fluorescent protein (GFP) and a streptavidin-binding peptide (SBP) due to the interaction with a molar excess of streptavidin molecules ("hooks") targeted to selected subcellular compartments. Supplementation of biotin competitively disrupts the interaction between the SBP moiety and streptavidin, liberating the chimeric target protein from its hooks, while addition of avidin causes the removal of biotin from the system and reestablishes the interaction. Based on this principle, we engineered two chimeric proteins involved in autophagy, namely microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B, best known as LC3) and sequestosome-1 (SQSTM1, best known as p62) to move them as SBP-GFP-LC3 and p62-SBP-GFP at will between the cytosol and two different organelles, the endoplasmic reticulum (ER) and the Golgi apparatus. Although both proteins were functional in thus far that SBP-GFP-LC3 and p62-SBP-GFP could recruit their endogenous binding partners, p62 and LC3, respectively, their enforced relocation to the ER or Golgi failed to induce organelle-specific autophagy. Hence, artificial tethering of LC3 or p62 to the surface of the ER and the Golgi is not sufficient to trigger autophagy.
Identifiants
pubmed: 31601788
doi: 10.1038/s41419-019-2011-5
pii: 10.1038/s41419-019-2011-5
pmc: PMC6787181
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
MAP1LC3A protein, human
0
Microtubule-Associated Proteins
0
P62 protein, human
0
RNA-Binding Proteins
0
Recombinant Proteins
0
Green Fluorescent Proteins
147336-22-9
Biotin
6SO6U10H04
Streptavidin
9013-20-1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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