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E3 ligase SCFSKP2 ubiquitinates and degrades tumor suppressor C/EBPα in acute myeloid leukemia.

Life sciences (2020-07-06)
Gatha Thacker, Mukul Mishra, Akshay Sharma, Anil Kumar Singh, Sabyasachi Sanyal, Arun Kumar Trivedi
RESUMEN

Transcription factor CCAAT/Enhancer binding protein alpha (C/EBPα) is a key regulator of myeloid differentiation, granulopoiesis in particular. Although CEBPA mutations are found in more than 10% in AML, functional inhibition of C/EBPα protein is also widely observed in AML. Here, we sought to examine if SKP2, an aberrantly enhanced E3 ubiquitin ligase in primary AMLs inhibits C/EBPα stability to induce differentiation block. Here we employed cell based assays such as transfections, immunoblotting, co-immunoprecipitation, luciferase and gel shift assays along with differentiation assays to investigate SKP2 regulated C/EBPα protein stability in acute myeloid leukemia. Here we discovered that oncogenic E3 ubiquitin ligase SCFskp2 ubiquitinates and destabilizes C/EBPα in a proteasome-dependent manner. Our data demonstrates that SKP2 physically interacts with C-terminal of C/EBPα and promotes its K48-linked ubiquitination-mediated degradation leading to its reduced transactivation potential, DNA binding ability and cellular functions. We further show that while overexpression of SKP2 inhibits both ectopic as well as endogenous C/EBPα in heterologous (HEK293T) as well as myeloid leukemia cells respectively, SKP2 depletion restores endogenous C/EBPα leading to reduced colony formation and enhanced myeloid differentiation of myeloid leukemia cells. Using Estradiol-inducible K562-C/EBPα-ER cells as yet another model of granulocytic differentiation, we further confirmed that SKP2 overexpression indeed inhibits granulocytic differentiation by mitigating C/EBPα stability. Our findings identify SKP2 as a potential negative regulator of C/EBPα stability and function in AML which suggests that SKP2 can be potentially targeted in AML to restore C/EBPα and overcome differentiation block.

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MISSION® esiRNA, targeting human SKP2
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Número de referencia del producto (SKU)Tamaño de envaseDisponibilidadPrecioCantidad
20 μg
Normalmente se envían en 3 semanas (de 4 a 6 semanas para pedidos personalizados).
$220.00
50 μg
Normalmente se envían en 3 semanas (de 4 a 6 semanas para pedidos personalizados).
$391.00