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Min Deng1, Fahui Li1, Bryan A. Ballif?2, Shan Li||, Xi Chen**, Lin Guo**, and Xin Ye3
From the Department of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China, the ?Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, the ||Shandong Normal University, Jinan, Shandong 250014, China, the **College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China, and the Graduate University of Chinese Academy of Sciences, Beijing 100101, China
Cyclin E/Cdk2 is a key regulator in G1-S transition. We have identified a novel cyclin E/Cdk2 substrate called Ankrd17 (ankyrin repeat protein 17) using the TAP tag purification technique. Ankrd17 protein contains two clusters of a total 25 ankyrin repeats at its N terminus, one NES (nuclear exporting signal) and one NLS (nuclear localization signal) in the middle, and one RXL motif at its C terminus. Ankrd17 is expressed in various tissues and associates with cyclin E/Cdk2 in an RXL-dependent manner. It can be phosphorylated by cyclin E/Cdk2 at 3 phosphorylation sites (Ser1791, Ser1794, and Ser2150). Overexpression of Ankrd17 promotes S phase entry, whereas depletion of Ankrd17 expression by small interfering RNA inhibits DNA replication and blocks cell cycle progression as well as up-regulates the expression of p53 and p21. Ankrd17 is localized to the nucleus and interacts with DNA replication factors including MCM family members, Cdc6 and PCNA. Depletion of Ankrd17 results in decreased loading of Cdc6 and PCNA onto DNA suggesting that Ankrd17 may be directly involved in the DNA replication process. Taken together, these data indicate that Ankrd17 is an important downstream effector of cyclin E/Cdk2 and positively regulates G1/S transition.
