Perinuclear positioning of endosomes can affect PS-ASO activities.
Animals
Biological Transport
Cell Line, Tumor
Cell Nucleus
/ metabolism
Cells, Cultured
Dyneins
/ metabolism
Endoplasmic Reticulum
/ metabolism
Endosomes
/ metabolism
HeLa Cells
Humans
Mice
Microscopy, Confocal
Microtubules
/ metabolism
Motion
Neoplasm Proteins
/ metabolism
Oligonucleotides, Antisense
/ genetics
Thionucleotides
/ genetics
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
16 12 2021
16 12 2021
Historique:
accepted:
22
11
2021
revised:
16
11
2021
received:
24
08
2021
pubmed:
9
12
2021
medline:
11
1
2022
entrez:
8
12
2021
Statut:
ppublish
Résumé
Phosphorothioate (PS) modified antisense oligonucleotide (ASO) drugs that act on cellular RNAs must enter cells and be released from endocytic organelles to elicit antisense activity. It has been shown that PS-ASOs are mainly released by late endosomes. However, it is unclear how endosome movement in cells contributes to PS-ASO activity. Here, we show that PS-ASOs in early endosomes display Brownian type motion and migrate only short distances, whereas PS-ASOs in late endosomes (LEs) move linearly along microtubules with substantial distances. In cells with normal microtubules and LE movement, PS-ASO-loaded LEs tend to congregate perinuclearly. Disruption of perinuclear positioning of LEs by reduction of dynein 1 decreased PS-ASO activity, without affecting PS-ASO cellular uptake. Similarly, disruption of perinuclear positioning of PS-ASO-LE foci by reduction of ER tethering proteins RNF26, SQSTM1 and UBE2J1, or by overexpression of P50 all decreased PS-ASO activity. However, enhancing perinuclear positioning through reduction of USP15 or over-expression of RNF26 modestly increased PS-ASO activity, indicating that LE perinuclear positioning is required for ensuring efficient PS-ASO release. Together, these observations suggest that LE movement along microtubules and perinuclear positioning affect PS-ASO productive release.
Identifiants
pubmed: 34878127
pii: 6456228
doi: 10.1093/nar/gkab1198
pmc: PMC8682747
doi:
Substances chimiques
Neoplasm Proteins
0
Oligonucleotides, Antisense
0
RNF26 protein, human
0
Thionucleotides
0
Dyneins
EC 3.6.4.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
12970-12985Informations de copyright
© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.
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