Quadruple and Truncated MEK3 Mutants Identified from Acute Lymphoblastic Leukemia Promote Degradation and Enhance Proliferation
Abstract
:1. Introduction
2. Results
2.1. Identification of Potential Oncogenes Involved in ALL
2.2. Identification of MEK3 Variants in ALL Patient Samples
2.3. Quadruple and Truncated MEK3 Mutants Decrease Protein Stability
2.4. Quadruple MEK3 Mutant Was Degraded at an Accelerated Rate Compared to Wild-Type Protein
2.5. Cysteine/Serine Protease Inhibitors Rescue Quadruple MEK3 Mutant Protein Expression
2.6. Proteosomal Degradation of Truncated MEK3 Mutant Is Ubiquitin Independent
2.7. MEK3 Mutants Disrupt Auto-Phosphorylation at T222 and Reduce T180/Y182 Phosphorylation of p38 MAPK
2.8. Quadruple and Truncated MEK3 Mutants Exhibited Less Suppresive Activity on the Viability of HEK293 Cells
3. Discussion
4. Materials and Methods
4.1. Whole-exome sequencing
4.2. Bioinformatics
4.3. Cell Culture
4.4. Plasmid and Site-Directed Mutagenesis
4.5. Transfection, Lysis of Cells, Immunoprecipitation, and Western Blots
4.6. Immunofluorescence
4.7. Protein Half-Life Studies
4.8. Cell Viability Assay
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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MAP2K3 | Provean | A.A.From | Location | A.A.To | Patients | Controls |
---|---|---|---|---|---|---|
chr17.21202191.c.a.c | −5.539 | P | 11 | T | 9 | 0 |
chr17.21202237.g.c.g | −2.605 | R | 26 | T | 9 | 0 |
chr17.21204187.g.t.g | −6.557 | R | 65 | L | 9 | 0 |
chr17.21204192.c.t.c | −5.085 | R | 67 | W | 9 | 0 |
chr17.21204210.c.t.c | −7.739 | Q | 73 | * | 8 | 0 |
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Ayala-Marin, Y.M.; Grant, A.H.; Rodriguez, G.; Kirken, R.A. Quadruple and Truncated MEK3 Mutants Identified from Acute Lymphoblastic Leukemia Promote Degradation and Enhance Proliferation. Int. J. Mol. Sci. 2021, 22, 12210. https://doi.org/10.3390/ijms222212210
Ayala-Marin YM, Grant AH, Rodriguez G, Kirken RA. Quadruple and Truncated MEK3 Mutants Identified from Acute Lymphoblastic Leukemia Promote Degradation and Enhance Proliferation. International Journal of Molecular Sciences. 2021; 22(22):12210. https://doi.org/10.3390/ijms222212210
Chicago/Turabian StyleAyala-Marin, Yoshira M., Alice H. Grant, Georgialina Rodriguez, and Robert A. Kirken. 2021. "Quadruple and Truncated MEK3 Mutants Identified from Acute Lymphoblastic Leukemia Promote Degradation and Enhance Proliferation" International Journal of Molecular Sciences 22, no. 22: 12210. https://doi.org/10.3390/ijms222212210