Yoon Hyun-Joo, Feoktistova Anna, Chen Jun-Song, Jennings Jennifer L, Link Andrew J, Gould Kathleen L
Howard Hughes Medical Institute and the Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
J Biol Chem. 2006 Oct 27;281(43):32284-93. doi: 10.1074/jbc.M603867200. Epub 2006 Sep 1.
The anaphase-promoting complex/cyclosome (APC/C) is a conserved multisubunit ubiquitin ligase required for the degradation of key cell cycle regulators. The APC/C becomes active at the metaphase/anaphase transition and remains active during G(1) phase. One mechanism linked to activation of the APC/C is phosphorylation. Although many sites of mitotic phosphorylation have been identified in core components of the APC/C, the consequence of any individual phosphorylation event has not been elucidated in vivo. In this study, we show that Hcn1 is an essential core component of the fission yeast APC/C and is critical for maintaining complex integrity. Moreover, Hcn1 is a phosphoprotein in vivo. Phosphorylation of Hcn1 occurs at a single Cdk1 site in vitro and in vivo. Mutation of this site to alanine, but not aspartic acid, compromises APC/C function and leads to a specific defect in the completion of cell division.
后期促进复合物/细胞周期体(APC/C)是一种保守的多亚基泛素连接酶,是降解关键细胞周期调节因子所必需的。APC/C在中期/后期转换时激活,并在G1期保持活性。与APC/C激活相关的一种机制是磷酸化。尽管在APC/C的核心组分中已鉴定出许多有丝分裂磷酸化位点,但任何单个磷酸化事件在体内的后果尚未阐明。在本研究中,我们表明Hcn1是裂殖酵母APC/C的必需核心组分,对维持复合物完整性至关重要。此外,Hcn1在体内是一种磷蛋白。Hcn1的磷酸化在体外和体内均发生在单个Cdk1位点。该位点突变为丙氨酸而非天冬氨酸会损害APC/C功能,并导致细胞分裂完成时出现特定缺陷。