Novel Insights into the Biochemical Mechanism of CK1ε and its Functional Interplay with DDX3X

Abstract

Casein Kinase 1 epsilon (CK1ε) is a member of the serine (Ser)/threonine (Thr) CK1 family, known to have crucial roles in several biological scenarios and, ever more frequently, in pathological contexts, such as cancer. Recently, the human DEAD-box RNA helicase 3 X-linked (DDX3X), involved in cancer proliferation and viral infections, has been identified as one of CK1ε substrates and its positive regulator in the Wnt/β-catenin network. However, the way by which these two proteins influence each other has not been fully clarified. In order to further investigate their interplay, we defined the kinetic parameters of CK1ε towards its substrates: ATP, casein, Dvl2 and DDX3X. CK1ε affinity for ATP depends on the nature of the substrate: increasing of casein concentrations led to an increase of KmATP, while increasing DDX3X reduced it. In literature, DDX3X is described to act as an allosteric activator of CK1ε. However, when we performed kinase reactions combining DDX3X and casein, we did not find a positive effect of DDX3X on casein phosphorylation by CK1ε, while both substrates were phosphorylated in a competitive manner. Moreover, CK1ε positively stimulates DDX3X ATPase activity. Our data provide a more detailed kinetic characterization on the functional interplay of these two proteins.

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“…Dvl2 phosphorylation by Ck1ε is dependent on its association with DDX3 (considered as a regulatory subunit of Ck1ε) (51). Ck1ε has also been reported to bind to ATP-dependent RNA helicase DDX3X allosterically, activating Wnt signaling (52). On the other hand, AMP-activated Protein kinase (AMPK) phosphorylates Ser389 of Ck1ε and upregulates its kinase activity (53).…”