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EVI1 oncoprotein expression and CtBP1-association oscillate through the cell cycle.

Molecular biology reports (2020-09-27)
Roberto Paredes, Marion Schneider, Stella Pearson, Hsiang Yin Teng, James R Kelly, Andrew Pierce, Tim C P Somervaille, Anthony D Whetton, Stefan Meyer

Aberrantly high expression of EVI1 in acute myeloid leukaemia (AML) is associated with poor prognosis. For targeted treatment of EVI1 overexpressing AML a more detailed understanding of aspects of spatiotemporal interaction dynamics of the EVI1 protein is important. EVI1 overexpressing SB1690CB AML cells were used for quantification and protein interaction studies of EVI1 and ΔEVI1. Cells were cell cycle-synchronised by mimosine and nocodazole treatment and expression of EVI1 and related proteins assessed by western blot, immunoprecipitation and immunofluorescence. EVI1 protein levels oscillate through the cell cycle, and EVI1 is degraded partly by the proteasome complex. Both EVI1 and ΔEVI1 interact with the co-repressor CtBP1 but dissociate from CtBP1 complexes during mitosis. Furthermore, a large fraction of EVI1, but not ΔEVI1 or CtBP1, resides in the nuclear matrix. In conclusion, EVI1- protein levels and EVI1-CtBP1 interaction dynamics vary though the cell cycle and differ between EVI1 and ΔEVI1. These data ad to the functional characterisation of the EVI1 protein in AML and will be important for the development of targeted therapeutic approaches for EVI1-driven AML.

Product Number
Product Description

Triton X-100, laboratory grade
TWEEN® 20, viscous liquid
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Phosphatase Inhibitor Cocktail 3, DMSO solution
Digitonin, Used as non-ionic detergent
n-Dodecyl β-D-maltoside, ≥98% (GC)
(±)-cis-4,4′-Dimethylaminorex solution, 1.0 mg/mL (1% 1M HCl in Methanol), ampule of 1 mL, certified reference material, Cerilliant®
Nocodazole, ≥99% (TLC), powder
BICINE, ≥99% (titration)
CHAPS hydrate, BioReagent, suitable for electrophoresis, ≥98% (HPLC)