Institute of Molecular Biology gGmbH (IMB), Ackermannweg 4, 55128 Mainz, Germany.
Université de Strasbourg, UMR7242 Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie de Strasbourg, 10413 Illkirch, Strasbourg, France.
Mol Cell. 2022 Apr 21;82(8):1589-1602.e5. doi: 10.1016/j.molcel.2022.02.016. Epub 2022 Mar 8.
A polyubiquitin chain can adopt a variety of shapes, depending on how the ubiquitin monomers are joined. However, the relevance of linkage for the signaling functions of polyubiquitin chains is often poorly understood because of our inability to control or manipulate this parameter in vivo. Here, we present a strategy for reprogramming polyubiquitin chain linkage by means of tailor-made, linkage- and substrate-selective ubiquitin ligases. Using the polyubiquitylation of the budding yeast replication factor PCNA in response to DNA damage as a model case, we show that altering the features of a polyubiquitin chain in vivo can change the fate of the modified substrate. We also provide evidence for redundancy between distinct but structurally similar linkages, and we demonstrate by proof-of-principle experiments that the method can be generalized to targets beyond PCNA. Our study illustrates a promising approach toward the in vivo analysis of polyubiquitin signaling.
多聚泛素链可以采用多种形状，具体取决于泛素单体的连接方式。然而，由于我们无法在体内控制或操纵这一参数，因此多聚泛素链连接对于泛素信号传递功能的相关性通常理解得很差。在这里，我们提出了一种通过定制的、连接和底物选择性的泛素连接酶来重新编程多聚泛素链连接的策略。我们以酵母复制因子 PCNA 在 DNA 损伤响应时的多泛素化作为模型案例，表明在体内改变多泛素链的特征可以改变修饰底物的命运。我们还提供了不同但结构相似的连接之间存在冗余的证据，并通过原理验证实验证明该方法可以推广到除 PCNA 之外的靶标。我们的研究说明了一种有前途的方法，可以对体内的泛素信号传递进行分析。
