Sun Yue, Gaio Daniela, Xie Bokun, Noma Kentaro, Wu Zilu, Jin Yishi
University of California, San Diego, La Jolla, California, United States.
MicroPubl Biol. 2024 Dec 5;2024. doi: 10.17912/micropub.biology.001396. eCollection 2024.
The E3 ubiquitin ligase RPM-1 consists of 3,766 amino acids, with a RING finger domain at the C-terminus that functions to target the DLK-1 kinase for degradation for synapse development and axon termination. for aka F07B7.12 resides 35 kb away from on chromosome V, and is a near-perfect 12 kb duplication of including the entire promoter region and coding sequences. RPMS-1 consists of 1,964 amino acids and is identical to the N-terminal half of RPM-1 , except the last 40 amino acids. Previous studies showed that transgenic overexpression of the duplicated region of did not rescue synapse defects of loss of function mutants. Here, using CRISPR editing, we generated a double knockout of and . We find that axon and synapse defects in double mutants resemble those in single mutants. Expression levels of endogenously tagged DLK-1 protein are increased to a comparable degree in and mutants, compared to the control. These data, along with previous transgene expression analysis, support the idea that does not have a major role in RPM-1-mediated cellular processes.
E3泛素连接酶RPM-1由3766个氨基酸组成，在C端有一个环状结构域，其作用是靶向DLK-1激酶进行降解，以促进突触发育和轴突终止。aka F07B7.12位于五号染色体上距离其35 kb处，是一个近乎完美的12 kb重复序列，包括整个启动子区域和编码序列。RPMS-1由1964个氨基酸组成，与RPM-1的N端一半相同，但最后40个氨基酸除外。先前的研究表明，重复区域的转基因过表达并不能挽救功能缺失突变体的突触缺陷。在这里，我们使用CRISPR编辑技术产生了 和 的双敲除。我们发现 双突变体中的轴突和突触缺陷与 单突变体中的相似。与对照相比，内源性标记的DLK-1蛋白在 和 突变体中的表达水平升高到了可比程度。这些数据，连同先前的转基因表达分析，支持了 在RPM-1介导的细胞过程中没有主要作用这一观点。
