%0 Journal Article
%T Cyclin-dependent kinase 1-mediated phosphorylation of SET at serine 7 is essential for its oncogenic activity
%A Hong Shen
%A Jixin Dong
%A Ling Yin
%A Yi Xiao
%A Yongji Zeng
%A Yuanhong Chen
%J Archive of "Cell Death & Disease".
%D 2019
%R 10.1038/s41419-019-1621-2
%X a HeLa cells were treated with DMSO (control), taxol (100£¿nM for 16£¿h), or nocodazole (Noco, 100£¿ng/ml for 16£¿h). Total cell lysates were electrophoresed on regular and Phos-tag SDS polyacrylamide gels and probed with the indicated antibodies. Increased cyclin B1 levels marks cells in mitosis. An asterisk (*) marks the phosphorylated/shifted band. b HeLa cells were treated with nocodazole as indicated and cell lysates were further treated with (+) or without (£¿) ¦Ë phosphatase (ppase). Total cell lysates were probed with the indicated antibodies. Increased cyclin B1 levels marks cells in mitosis. An asterisk marks the phosphorylated/shifted band. c HeLa cells were treated with nocodazole, with or without various kinase inhibitors as indicated. Inhibitors were added 1.5£¿h before harvesting the cells (with MG132 to prevent cyclin B degradation and subsequent mitotic exit). The concentrations used for each inhibitor were as follows: VX680 2£¿¦ÌM, RO3306 5£¿¦ÌM, Purvalanol A 10£¿¦ÌM, BI-2536 100£¿nM, SB216763 10£¿¦ÌM, MK-2206 10£¿¦ÌM, SB203580 10£¿¦ÌM, U0126 20£¿¦ÌM, SP600125 20£¿¦ÌM, {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002 30£¿¦ÌM, and rapamycin 100£¿nM. Total cell lysates were electrophoresed on regular and Phos-tag SDS polyacrylamide gels and probed with the indicated antibodies. Increased cyclin B1 levels mark cells in mitosis. An asterisk marks the phosphorylated/shifted band. d In vitro kinase assays with purified CDK1/cyclin B1 complex using GST-tagged SET isoform 1 proteins as substrates. RO3306 (5£¿¦ÌM) was used to inhibit CDK1/cyclin B1 kinase activity. e GST-SET and GST-SET-S7A proteins were used for in vitro kinase assays with purified CDK1/cyclin B1 complex. f In vitro kinase assays were done as in e except anti-phospho-SET S7 antibody was use
%K Cell growth
%K Cell signalling
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6522553/