Set Protein Is Involved in FLT3 Membrane Trafficking
Abstract
:Simple Summary
Abstract
1. Introduction
2. Material and Methods
2.1. General Methodology
2.2. Cell Culture and Treatments
2.3. Immunofluorescence
2.4. Fluorescence-Activated Cell Sorting (FACS)
2.5. RNA Immunoprecipitation (RIP)
2.6. siRNA Transfection
2.7. Ectopic Expression of FLT3-WT and FLT3-ITD in Ba/F3 Cells
2.8. Statistical Analysis
3. Results
3.1. FLT3 Interacts with SET in FLT3-WT Cells, but Not in FLT3-ITD Cells
3.2. FLT3-WT mRNA Forms a Complex with HuR and SET
3.3. SET Modulates FLT3-WT Membrane Transport
3.4. SET Binds to FLT3 Underglycosylated in the ER
3.5. FLT3 Inactivation Enhances SET/FLT3 Binding in FLT3-ITD Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
AML | acute myeloid leukemia |
AREs | AU-rich elements |
CK2 | casein kinase 2 |
ER | endoplasmic reticulum |
FLT3 | Fms-related tyrosine kinase 3 |
FLT3LG | Fms-related tyrosine kinase 3-ligand |
ITD | internal tandem duplication |
PP2A | protein phosphatase 2A |
RIP | RNA immunoprecipitation |
TKI | tyrosine kinase inhibitors |
UPR | unfolded protein response |
WB | western blot |
WT | wild type |
References
- Griffith, J.; Black, J.; Faerman, C.; Swenson, L.; Wynn, M.; Lu, F.; Lippke, J.; Saxena, K. The structural basis for autoinhibition of FLT3 by the juxtamembrane domain. Mol. Cell 2004, 13, 169–178. [Google Scholar] [CrossRef] [PubMed]
- Heiss, E.; Masson, K.; Sundberg, C.; Pedersen, M.; Sun, J.; Bengtsson, S.; Ronnstrand, L. Identification of Y589 and Y599 in the juxtamembrane domain of Flt3 as ligand-induced autophosphorylation sites involved in binding of Src family kinases and the protein tyrosine phosphatase SHP2. Blood 2006, 108, 1542–1550. [Google Scholar] [CrossRef] [PubMed]
- Walter, M.; Lucet, I.S.; Patel, O.; Broughton, S.E.; Bamert, R.; Williams, N.K.; Fantino, E.; Wilks, A.F.; Rossjohn, J. The 2.7 A crystal structure of the autoinhibited human c-Fms kinase domain. J. Mol. Biol. 2007, 367, 839–847. [Google Scholar] [CrossRef] [PubMed]
- Stirewalt, D.L.; Kopecky, K.J.; Meshinchi, S.; Engel, J.H.; Pogosova-Agadjanyan, E.L.; Linsley, J.; Slovak, M.L.; Willman, C.L.; Radich, J.P. Size of FLT3 internal tandem duplication has prognostic significance in patients with acute myeloid leukemia. Blood 2006, 107, 3724–3726. [Google Scholar] [CrossRef] [Green Version]
- Schnittger, S.; Bacher, U.; Haferlach, C.; Alpermann, T.; Kern, W.; Haferlach, T. Diversity of the juxtamembrane and TKD1 mutations (exons 13–15) in the FLT3 gene with regards to mutant load, sequence, length, localization, and correlation with biological data. Genes Chromosomes Cancer 2012, 51, 910–924. [Google Scholar] [CrossRef]
- Choudhary, C.; Schwable, J.; Brandts, C.; Tickenbrock, L.; Sargin, B.; Kindler, T.; Fischer, T.; Berdel, W.E.; Muller-Tidow, C.; Serve, H. AML-associated Flt3 kinase domain mutations show signal transduction differences compared with Flt3 ITD mutations. Blood 2005, 106, 265–273. [Google Scholar] [CrossRef]
- Kantarjian, H.; Kadia, T.; DiNardo, C.; Daver, N.; Borthakur, G.; Jabbour, E.; Garcia-Manero, G.; Konopleva, M.; Ravandi, F. Acute myeloid leukemia: Current progress and future directions. Blood Cancer J. 2021, 11, 41. [Google Scholar] [CrossRef]
- Döhner, H.; Wei, A.H.; Löwenberg, B. Towards precision medicine for AML. Nat. Rev. Clin. Oncol. 2021, 18, 577–590. [Google Scholar] [CrossRef]
- Perl, A.E. Availability of FLT3 inhibitors: How do we use them? Blood 2019, 134, 741–745. [Google Scholar] [CrossRef]
- Smith, C.C. The growing landscape of FLT3 inhibition in AML. Hematol. Am. Soc. Hematol. Educ. Program 2019, 2019, 539–547. [Google Scholar] [CrossRef]
- Döhner, H.; Wei, A.H.; Appelbaum, F.R.; Craddock, C.; DiNardo, C.D.; Dombret, H.; Ebert, B.L.; Fenaux, P.; Godley, L.A.; Hasserjian, R.P.; et al. Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN. Blood 2022, 140, 1345–1377. [Google Scholar] [CrossRef]
- Cristóbal, I.; Garcia-Orti, L.; Cirauqui, C.; Cortes-Lavaud, X.; García-Sánchez, M.A.; Calasanz, M.J.; Odero, M.D. Overexpression of SET is a recurrent event associated with poor outcome and contributes to protein phosphatase 2A inhibition in acute myeloid leukemia. Haematologica 2012, 97, 543–550. [Google Scholar] [CrossRef] [Green Version]
- Yu, G.; Yan, T.; Feng, Y.; Liu, X.; Xia, Y.; Luo, H.; Wang, J.Z.; Wang, X. Ser9 phosphorylation causes cytoplasmic detention of I2PP2A/SET in Alzheimer disease. Neurobiol. Aging 2013, 34, 1748–1758. [Google Scholar] [CrossRef]
- Ten Klooster, J.P.; Leeuwen, I.V.; Scheres, N.; Anthony, E.C.; Hordijk, P.L. Rac1-induced cell migration requires membrane recruitment of the nuclear oncogene SET. EMBO J. 2007, 26, 336–345. [Google Scholar] [CrossRef] [Green Version]
- Fan, Z.; Beresford, P.J.; Oh, D.Y.; Zhang, D.; Lieberman, J. Tumor suppressor NM23-H1 is a granzyme A-activated DNase during CTL-mediated apoptosis, and the nucleosome assembly protein SET is its inhibitor. Cell 2003, 112, 659–672. [Google Scholar] [CrossRef] [Green Version]
- Arriazu, E.; Vicente, C.; Pippa, R.; Peris, I.; Martínez-Balsalobre, E.; García-Ramírez, P.; Marcotegui, N.; Igea, A.; Alignani, D.; Rifón, J.; et al. A new regulatory mechanism of protein phosphatase 2A activity via SET in acute myeloid leukemia. Blood Cancer J. 2020, 10, 3. [Google Scholar] [CrossRef] [Green Version]
- Mayr, C. Regulation by 3′-Untranslated Regions. Annu. Rev. Genet. 2017, 51, 171–194. [Google Scholar] [CrossRef] [Green Version]
- Berkovits, B.D.; Mayr, C. Alternative 3′UTRs act as scaffolds to regulate membrane protein localization. Nature 2015, 522, 363–367. [Google Scholar] [CrossRef] [Green Version]
- Ma, W.; Mayr, C. A Membraneless Organelle Associated with the Endoplasmic Reticulum Enables 3′UTR-Mediated Protein-Protein Interactions. Cell 2018, 175, 1492–1506. [Google Scholar] [CrossRef] [Green Version]
- Schindelin, J.; Arganda-Carreras, I.; Frise, E.; Kaynig, V.; Longair, M.; Pietzsch, T.; Preibisch, S.; Rueden, C.; Saalfeld, S.; Schmid, B.; et al. Fiji: An open-source platform for biological-image analysis. Nat. Methods 2012, 9, 676–682. [Google Scholar] [CrossRef] [Green Version]
- Schmidt-Arras, D.E.; Bohmer, A.; Markova, B.; Choudhary, C.; Serve, H.; Bohmer, F.D. Tyrosine phosphorylation regulates maturation of receptor tyrosine kinases. Mol. Cell. Biol. 2005, 25, 3690–3703. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schmidt-Arras, D.; Böhmer, S.A.; Koch, S.; Müller, J.P.; Blei, L.; Cornils, H.; Bauer, R.; Korasikha, S.; Thiede, C.; Böhmer, F.D. Anchoring of FLT3 in the endoplasmic reticulum alters signaling quality. Blood 2009, 113, 3568–3576. [Google Scholar] [CrossRef] [PubMed]
- Reiter, K.; Polzer, H.; Krupka, C.; Maiser, A.; Vick, B.; Rothenberg-Thurley, M.; Metzeler, K.H.; Dörfel, D.; Salih, H.R.; Jung, G.; et al. Tyrosine kinase inhibition increases the cell surface localization of FLT3-ITD and enhances FLT3-directed immunotherapy of acute myeloid leukemia. Leukemia 2018, 32, 313–322. [Google Scholar] [CrossRef] [Green Version]
- DiNardo, C.D.; Wei, A.H. How I treat acute myeloid leukemia in the era of new drugs. Blood 2020, 135, 85–96. [Google Scholar] [CrossRef] [PubMed]
- Mayr, C. What Are 3′UTRs Doing? Cold Spring Harb. Perspect. Biol. 2019, 11, a034728. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lee, S.H.; Mayr, C. Gain of Additional BIRC3 Protein Functions through 3′-UTR-Mediated Protein Complex Formation. Mol. Cell 2019, 74, 701–712. [Google Scholar] [CrossRef]
- Lv, K.; Ren, J.G.; Han, X.; Gui, J.; Gong, C.; Tong, W. Depalmitoylation rewires FLT3-ITD signaling and exacerbates leukemia progression. Blood 2021, 138, 2244–2255. [Google Scholar] [CrossRef]
- He, X.; Zhu, Y.; Lin, Y.C.; Li, M.; Du, J.; Dong, H.; Sun, J.; Zhu, L.; Wang, H.; Ding, Z.; et al. PRMT1-mediated FLT3 arginine methylation promotes maintenance of FLT3-ITD+ acute myeloid leukemia. Blood 2019, 134, 548–560. [Google Scholar] [CrossRef]
- Duan, C.; Fukuda, T.; Isaji, T.; Qi, F.; Yang, J.; Wang, Y.; Takahashi, S.; Gu, J. Deficiency of core fucosylation activates cellular signaling dependent on FLT3 expression in a Ba/F3 cell system. FASEB J. 2020, 34, 3239–3252. [Google Scholar] [CrossRef]
- Hu, X.; Chen, F. Targeting on glycosylation of mutant FLT3 in acute myeloid leukemia. Hematology 2019, 24, 651–660. [Google Scholar] [CrossRef] [Green Version]
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Share and Cite
Marcotegui, N.; Romero-Murillo, S.; Marco-Sanz, J.; Peris, I.; Berrozpe, B.S.; Vicente, C.; Odero, M.D.; Arriazu, E. Set Protein Is Involved in FLT3 Membrane Trafficking. Cancers 2023, 15, 2233. https://doi.org/10.3390/cancers15082233
Marcotegui N, Romero-Murillo S, Marco-Sanz J, Peris I, Berrozpe BS, Vicente C, Odero MD, Arriazu E. Set Protein Is Involved in FLT3 Membrane Trafficking. Cancers. 2023; 15(8):2233. https://doi.org/10.3390/cancers15082233
Chicago/Turabian StyleMarcotegui, Nerea, Silvia Romero-Murillo, Javier Marco-Sanz, Irene Peris, Blanca S. Berrozpe, Carmen Vicente, María D. Odero, and Elena Arriazu. 2023. "Set Protein Is Involved in FLT3 Membrane Trafficking" Cancers 15, no. 8: 2233. https://doi.org/10.3390/cancers15082233