CRY Drives Cyclic CK2-Mediated BMAL1 Phosphorylation to Control the Mammalian Circadian Clock
Fig 3
CRY is required for circadian CK2-mediated BMAL1-Ser90 phosphorylation, with inverse phase to BMAL1–CK2β binding rhythm.
BMAL1 phosphorylation and CK2β, CRY1, and CRY2 interaction profiles in wild type (WT), CRYdKO (Cry1-/-/Cry2-/-) MEFs (A) and CRYdKO MEFs stably expressing mCry1 promoter-driven Myc-CRY1 or CMV-promoter-driven Myc-GFP (control) (B). Cells were Dex-synchronized and harvested at 6 h intervals. BMAL1-IP and lysates were subjected to IB analysis for P-BMAL1-S90, CK2β, CRY1, CRY2 and actin (loading control). Shown are representative examples of n = 3 experiments (upper panels). P-BMAL1-S90 and BMAL1-bound CK2β, CRY1 and CRY2 levels were quantified and normalized against actin levels (lower panels). Maximum values in WT/CRYdKO+CRY1 were set as 1. Error bars indicate SD.