Pho85 cyclins (Pcls), activators of the yeast cyclin-dependent kinase (CDK) Pho85, belong together with the p35 activator of mammalian CDK5 to a distinct structural cyclin class. Different Pcls target Pho85 to distinct substrates. Pcl5 targets Pho85 specifically to Gcn4, a yeast transcription factor involved in the response to amino acid starvation, eventually causing the degradation of Gcn4. Pcl5 is itself highly unstable, an instability that was postulated to be important for regulation of Gcn4 degradation. We used hybrids between different Pcls to circumscribe the substrate recognition function to the core cyclin box domain of Pcl5. Furthermore, the cyclin hybrids revealed that Pcl5 degradation is uniquely dependent on two distinct degradation signals: one N-terminal and one C-terminal to the cyclin box domain. Whereas the C-terminal degradation signal is independent of Pho85, the N-terminal degradation signal requires phosphorylation of a specific threonine residue by the Pho85 molecule bound to the cyclin. This latter mode of degradation depends on the SCF ubiquitin ligase. Degradation of Pcl5 after self-catalyzed phosphorylation ensures that activity of the Pho85/Pcl5 complex is self-limiting in vivo. We demonstrate the importance of this mechanism for the regulation of Gcn4 degradation and for cell growth under conditions of amino acid starvation.

译文

:Pho85细胞周期蛋白(Pcls)是酵母细胞周期蛋白依赖性激酶(CDK)Pho85的激活剂,与哺乳动物CDK5的p35激活剂一起属于独特的结构细胞周期蛋白类别。不同的Pcls将Pho85靶向不同的底物。 Pcl5将Pho85特异性靶向Gcn4,Gcn4是参与对氨基酸饥饿反应的酵母转录因子,最终导致Gcn4降解。 Pcl5本身是高度不稳定的,据认为这种不稳定性对于调节Gcn4降解很重要。我们使用不同Pcls之间的杂合体将底物识别功能限制在Pcl5的核心细胞周期蛋白框结构域中。此外,细胞周期蛋白杂合体显示Pcl5降解唯一依赖于两个不同的降解信号:细胞周期蛋白盒结构域的一个N端和一个C端。尽管C末端降解信号独立于Pho85,但N末端降解信号需要结合到细胞周期蛋白上的Pho85分子使特定的苏氨酸残基磷酸化。后一种降解方式取决于SCF泛素连接酶。自催化磷酸化后Pcl5的降解可确保Pho85 / Pcl5复合物的活性在体内是自限性的。我们证明了这种机制对于调节Gcn4降解和氨基酸饥饿条件下细胞生长的重要性。

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