Fig. 4: The CDK1-cyclin B complex accounts for cGAS phosphorylation and inactivation in mitotic cells.
From: Phosphorylation of cGAS by CDK1 impairs self-DNA sensing in mitosis
a CDK1 but not AKT or CDK2 inhibitor abolishes mcGAS S291 phosphorylation. Raw264.7 cells were arrested at mitosis with nocodazole (Noc arrest) and then treated with the indicated kinase inhibitors (10 μM) for 15 min before immunoblotting or FACS analysis. b cGAS is a direct substrate of the CDK1-cyclin B complex. In vitro kinase assays were performed with recombinant CDK1-Cyclin B proteins and FLAG-cGAS immunoaffinity-purified from THP1 cells stably-expressing FLAG-cGAS in the presence or absence of RO-3306 (10 μM). The reactions were then analyzed by immunoblotting analysis with the indicated antibodies. c The CDK1-cylin B1 complex impairs cGAS activation. MITA stably-expressing HEK293 cells were transfected with the indicated plasmids for 24 h before luciferase assays. ***P < 0.001 (Student’s t-test, unpaired, two-tailed). Data shown are mean ± S.D. of one representative experiment performed in triplet. Data are representative of three biological repeats.
