Induction of ER Stress in Acute Lymphoblastic Leukemia Cells by the Deubiquitinase Inhibitor VLX1570
Abstract
:1. Introduction
2. Results
2.1. The DUB Inhibitor VLX1570 Elicits ER Stress in ALL Cell Lines, but only Weak Induction of CHOP
2.2. The eIF2B Activator ISRIB Enhances Induction of BiP
2.3. Bortezomib and VLX1570 Inhibit Translation
2.4. Limited Effects of the eIF2B Activator ISRIB on Translational Inhibition by VLX1570
2.5. Evidence for Elongational Block by VLX1570 Treatment
2.6. VLX1570 Reduces the Proliferation of RS4:11 Cells in Zebrafish Embryos
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.3. Protein Synthesis
4.4. Immunoblotting
4.5. Polysome Fractionation
4.6. Cell Labelling
4.7. Zebrafish Assay
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ALL | Acute lymphoblastic leukemia |
ATF4 | Activating Transcription Factor 4 |
CHOP | C/EBP homologous protein |
CML | Chronic myeloid leukemia |
CHX | Cyclohexamide |
DiI | 1,1′-dioctadecyl-3,3,3′3´-tetramethylindocarbocyanine |
DUB | Deubiquitinase |
ER | Endoplasmic reticulum |
eIF2α | Eukaryotic initiation factor 2α |
eIF2B | Eukaryotic translation initiation factor 2 subunit beta |
HRI | Heme-related eIF2α kinase |
HSP | Heat shock protein |
ISR | Integrated stress response |
ISRIB | Integrated Stress Response Inhibitor |
mTOR | Mammalian target of rapamycin |
PARP | Poly (ADP-ribose) polymerase |
PERK | Protein kinase R (PKR)-like endoplasmic reticulum kinase) |
TCA | Trichloroacetic acid |
UPR | Unfolded Protein Response |
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Pellegrini, P.; Selvaraju, K.; Faustini, E.; Mofers, A.; Zhang, X.; Ternerot, J.; Schubert, A.; Linder, S.; D′Arcy, P. Induction of ER Stress in Acute Lymphoblastic Leukemia Cells by the Deubiquitinase Inhibitor VLX1570. Int. J. Mol. Sci. 2020, 21, 4757. https://doi.org/10.3390/ijms21134757
Pellegrini P, Selvaraju K, Faustini E, Mofers A, Zhang X, Ternerot J, Schubert A, Linder S, D′Arcy P. Induction of ER Stress in Acute Lymphoblastic Leukemia Cells by the Deubiquitinase Inhibitor VLX1570. International Journal of Molecular Sciences. 2020; 21(13):4757. https://doi.org/10.3390/ijms21134757
Chicago/Turabian StylePellegrini, Paola, Karthik Selvaraju, Elena Faustini, Arjan Mofers, Xiaonan Zhang, Jens Ternerot, Alice Schubert, Stig Linder, and Pádraig D′Arcy. 2020. "Induction of ER Stress in Acute Lymphoblastic Leukemia Cells by the Deubiquitinase Inhibitor VLX1570" International Journal of Molecular Sciences 21, no. 13: 4757. https://doi.org/10.3390/ijms21134757