Dynamic ADP-ribosylation signaling is a crucial pathway that controls fundamental cellular processes, in particular, the response to cellular stresses such as DNA damage, reactive oxygen species, and infection. In some pathogenic microbes, the response to oxidative stress is controlled by a SirTM/zinc-containing macrodomain (Zn-Macro) pair responsible for establishment and removal of the modification, respectively. Targeting this defence mechanism against the host’s innate immune response may lead to novel approaches to support the fight against emerging antimicrobial resistance. Earlier studies suggested that Zn-Macros play a key role in the activation of this defence. Therefore, we used phylogenetic, biochemical, and structural approaches to elucidate the functional properties of these enzymes. Using the substrate mimetic asparagine-ADP-ribose as well as the ADP-ribose product, we characterize the catalytic role of the zinc ion in the removal of the ADP-ribosyl modification. Furthermore, we determined structural properties that contribute to substrate selectivity within the different Zn-Macro branches. Together, our data not only give new insights into the Zn-Macro family but also highlight their distinct features that may be exploited for the development of future therapies.

中文翻译：

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锌依赖性大结构域逆转 ADP-核糖基化的进化和分子基础


动态 ADP-核糖基化信号转导是控制基本细胞过程的关键途径，特别是对细胞应激（如 DNA 损伤、活性氧和感染）的反应。在一些病原微生物中，对氧化应激的反应由分别负责建立和去除修饰的 Sir TM /锌大结构域 （Zn-Macro） 对控制。针对宿主的先天免疫反应的这种防御机制可能会导致支持对抗新出现的抗菌素耐药性的新方法。早期研究表明，Zn-Macros 在激活这种防御中起着关键作用。因此，我们使用系统发育、生化和结构方法来阐明这些酶的功能特性。使用底物模拟天冬酰胺-ADP-核糖以及 ADP-核糖产物，我们表征了锌离子在去除 ADP-核糖基修饰中的催化作用。此外，我们确定了有助于不同 Zn-Macro 分支内底物选择性的结构特性。总之，我们的数据不仅为Zn-Macro家族提供了新的见解，还突出了它们的独特特征，这些特征可能被用于未来疗法的开发。