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通过非水解性无基化ADP-ribose识别RNF114作为ADPR-Ub读取器

Max S Kloet¹, Chatrin Chatrin², Rishov Mukhopadhyay¹

¹Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, the Netherlands.

Nature communications

|July 9, 2025

概括

研究人员探索了ubiquitination和ADP-ribosylation之间的相互作用,发现RNF114蛋白识别和修改ubiquitinatedADP-ribose,这对DNA损伤反应至关重要.

科学领域:

生物化学生物化学
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

翻译后的修改,如在细胞过程中的ubiquitination和ADP-ribosylation交联.
混合修饰,如ADP-ribose与ubiquitin相关,涉及细菌感染和DNA损伤反应.

研究的目的:

为了研究人类E3结合酶对ADP-ribose的泛化.
阐明 RNF114 蛋白在识别和处理这些混合变异中的作用.

主要方法:

准备一个不可水解的ADP-ribose-ubiquitin (ADPr-Ub) 探针.
蛋白质组学用于识别相互作用的蛋白质.
生物物理和生物化学测试以验证相互作用.
对RNF114.4的域删除分析.
激光诱导的DNA损伤测定.激光诱导的DNA损伤测定.

主要成果:

确定RNF114是一种与ADPR-Ub探测器相互作用的蛋白质.
RNF114优先结合于无基因的ADP-ribose,而不是未经修改的ubiquitin.
RNF114可以通过K11连接的ubiquitin链延长无化ADP-ribose.
RNF114的双联指 (ZnF2+ZnF3) 和随处相互作用动机 (UIM) 域对于识别随处的ADP-ribose至关重要.
这些RNF114域对于将蛋白质招募到DNA损伤部位至关重要.

结论:

RNF114在处理无处不在的ADP-ribose修饰中发挥着关键作用.
RNF114的特定域对于这种识别和随后的无处不在至关重要.
对于DNA损伤反应途径来说,RNF114在修改无素化ADP-ribose的功能非常重要.

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