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Protein kinase CK2 phosphorylates a conserved motif in the Notch effector E(spl)-Mγ

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Abstract

Across metazoan animals, the effects of Notch signaling are mediated via the Enhancer of Split (E(spl)/HES) basic Helix–Loop–Helix–Orange (bHLH-O) repressors. Although these repressors are generally conserved, their sequence diversity is, in large part, restricted to the C-terminal domain (CtD), which separates the Orange (O) domain from the penultimate WRPW tetrapeptide motif that binds the obligate co-repressor Groucho. While the kinases CK2 and MAPK target the CtD and regulate Drosophila E(spl)-M8 and mammalian HES6, the generality of this regulation to other E(spl)/HES repressors has remained unknown. To determine the broader impact of phosphorylation on this large family of repressors, we conducted bioinformatics, evolutionary, and biochemical analyses. Our studies identify E(spl)-Mγ as a new target of native CK2 purified from Drosophila embryos, reveal that phosphorylation is specific to CK2 and independent of the regulatory CK2-β subunit, and identify that the site of phosphorylation is juxtaposed to the WRPW motif, a feature unique to and conserved in the Mγ homologues over 50 × 106 years of Drosophila evolution. Thus, a preponderance of E(spl) homologues (four out of seven total) in Drosophila are targets for CK2, and the distinct positioning of the CK2 and MAPK sites raises the prospect that phosphorylation underlies functional diversity of bHLH-O proteins.

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All data generated or analyzed during this study are included in this published article.

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The authors thank Mohna Bandyopadhyay and Sophia Zhang for technical assistance.

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  1. Department of Biology Life Sciences Building, West Virginia University, 53 Campus Drive, Morgantown, WV, 26506-6057, USA

    Lucas M. Jozwick & Ashok P. Bidwai

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LJ and AB contributed to design of the work, as well as acquisition, analysis, and interpretation of data. LJ and AB drafted the work. LJ and AB approved the version to be published. LJ and AB agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

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Correspondence to Ashok P. Bidwai.

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Jozwick, L.M., Bidwai, A.P. Protein kinase CK2 phosphorylates a conserved motif in the Notch effector E(spl)-Mγ. Mol Cell Biochem 478, 781–790 (2023). https://doi.org/10.1007/s11010-022-04539-5

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