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Zou, Y, Mi, J, Wang, W, Lu, J, Zhao, W, Liu, Z, Hu, H, Yang, Y, Gao, X, Jiang, B, Shao, C and Gong, Y (2013) CUL4B promotes replication licensing by up-regulating the CDK2-CDC6 cascade. J. Cell Biol. 200:743-56
Cullin-RING ubiquitin ligases (CRLs) participate in the regulation of diverse cellular processes including cell cycle progression. Mutations in the X-linked CUL4B, a member of the cullin family, cause mental retardation and other developmental abnormalities in humans. Cells that are deficient in CUL4B are severely selected against in vivo in heterozygotes. Here we report a role of CUL4B in the regulation of replication licensing. Strikingly, CDC6, the licensing factor in replication, was positively regulated by CUL4B and contributed to the loading of MCM2 to chromatin. The positive regulation of CDC6 by CUL4B depends on CDK2, which phosphorylates CDC6, protecting it from APC(CDH1)-mediated degradation. Thus, aside being required for cell cycle reentry from quiescence, CDK2 also contributes to pre-replication complex assembly in G1 phase of cycling cells. Interestingly, the up-regulation of CDK2 by CUL4B is achieved via the repression of miR-372 and miR-373, which target CDK2. Our findings thus establish a CUL4B-CDK2-CDC6 cascade in the regulation of DNA replication licensing.
Cadherins/genetics; Cadherins/metabolism; Cell Cycle Proteins/genetics; Cell Cycle Proteins/metabolism; Chromatin/genetics; Chromatin/metabolism; Cullin Proteins/genetics; Cullin Proteins/metabolism; Cyclin-Dependent Kinase 2/genetics; Cyclin-Dependent Kinase 2/metabolism; DNA Replication/physiology; G1 Phase/physiology; HEK293 Cells; HeLa Cells; Humans; MicroRNAs/genetics; MicroRNAs/metabolism; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Up-Regulation/physiology