HSP70 regulates Eg5 distribution within the mitotic spindle and modulates the cytotoxicity of Eg5 inhibitors.
Antineoplastic Agents
/ metabolism
Cell Cycle Proteins
/ metabolism
Cell Proliferation
Drug Resistance, Neoplasm
/ physiology
HSP70 Heat-Shock Proteins
/ metabolism
HeLa Cells
Humans
Kinesins
/ metabolism
Microtubule-Associated Proteins
/ metabolism
Microtubules
/ metabolism
Mitosis
/ physiology
Spindle Apparatus
/ metabolism
Journal
Cell death & disease
ISSN: 2041-4889
Titre abrégé: Cell Death Dis
Pays: England
ID NLM: 101524092
Informations de publication
Date de publication:
01 09 2020
01 09 2020
Historique:
received:
13
04
2020
accepted:
07
08
2020
revised:
06
08
2020
entrez:
3
9
2020
pubmed:
3
9
2020
medline:
21
4
2021
Statut:
epublish
Résumé
The heat shock protein 70 (HSP70) is a conserved molecular chaperone and proteostasis regulator that protects cells from pharmacological stress and promotes drug resistance in cancer cells. In this study, we found that HSP70 may promote resistance to anticancer drugs that target the mitotic kinesin, Eg5, which is essential for assembly and maintenance of the mitotic spindle and cell proliferation. Our data show that loss of HSP70 activity enhances Eg5 inhibitor-induced cytotoxicity and spindle abnormalities. Furthermore, HSP70 colocalizes with Eg5 in the mitotic spindle, and inhibition of HSP70 disrupts this colocalization. Inhibition or depletion of HSP70 also causes Eg5 to accumulate at the spindle pole, altering microtubule dynamics and leading to chromosome misalignment. Using ground state depletion microscopy followed by individual molecule return (GSDIM), we found that HSP70 inhibition reduces the size of Eg5 ensembles and prevents their localization to the inter-polar region of the spindle. In addition, bis(maleimido)hexane-mediated protein-protein crosslinking and proximity ligation assays revealed that HSP70 inhibition deregulates the interaction between Eg5 tetramers and TPX2 at the spindle pole, leading to their accumulation in high-molecular-weight complexes. Finally, we showed that the passive substrate-binding activity of HSP70 is required for appropriate Eg5 distribution and function. Together, our results show that HSP70 substrate-binding activity may regulate proper assembly of Eg5 ensembles and Eg5-TPX2 complexes to modulate mitotic distribution/function of Eg5. Thus, HSP70 inhibition may sensitize cancer cells to Eg5 inhibitor-induced cytotoxicity.
Identifiants
pubmed: 32873777
doi: 10.1038/s41419-020-02919-7
pii: 10.1038/s41419-020-02919-7
pmc: PMC7462862
doi:
Substances chimiques
Antineoplastic Agents
0
Cell Cycle Proteins
0
HSP70 Heat-Shock Proteins
0
KIF11 protein, human
0
Microtubule-Associated Proteins
0
TPX2 protein, human
0
Kinesins
EC 3.6.4.4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
715Références
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