Yan Jing, Zhang Hengsen, Tomochika Yuna, Chen Bin, Ping Yashuang, Islam Md Shoriful, Aramaki Shuhei, Sato Tomohito, Nagashima Yu, Nakamura Tomohiko, Kahyo Tomoaki, Kaneda Daita, Ogawa Kenji, Yoshida Minoru, Setou Mitsutoshi
Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan.
Department of Radiation Oncology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan.
Biomedicines. 2023 Sep 8;11(9):2491. doi: 10.3390/biomedicines11092491.
Ubiquitin-like 3 (UBL3) is a membrane-anchored protein that plays a crucial role in sorting proteins into small extracellular vesicles. Aggregations of alpha-synuclein (α-syn) are associated with the pathology of neurodegenerative diseases such as Parkinson's disease. Recently, the interaction between UBL3 and α-syn was discovered, with potential implications in clearing excess α-syn from neurons and its role in disease spread. However, the regulator that can mediate the interaction between UBL3 and α-syn remains unclear. In this study, using the split gaussian luciferase complementation assay and RNA interference technology, we identified that QSOX2, HTATIP2, UBE3C, MGST3, NSF, HECTD1, SAE1, and ATG3 were involved in downregulating the interaction between UBL3 and α-syn. Notably, silencing MGST3 had the most significant impact. Immunocytochemistry staining confirmed the impact of MGST3 silencing on the co-localization of UBL3 and α-syn in cells. MGST3 is a part of the antioxidant system, and silencing MGST3 is believed to contribute to oxidative stress. We induced oxidative stress with hydrogen peroxide, observing its effect on the UBL3-α-syn interaction, and showing that 800 µM of HO downregulated this interaction. In conclusion, silencing MGST3 downregulates the interaction between UBL3 and α-syn.
泛素样蛋白3（UBL3）是一种膜锚定蛋白，在将蛋白质分选到小细胞外囊泡中发挥关键作用。α-突触核蛋白（α-syn）的聚集与帕金森病等神经退行性疾病的病理过程相关。最近，发现了UBL3与α-syn之间的相互作用，这对清除神经元中多余的α-syn及其在疾病传播中的作用具有潜在影响。然而，能够介导UBL3与α-syn之间相互作用的调节因子仍不清楚。在本研究中，我们使用分裂高斯荧光素酶互补分析和RNA干扰技术，确定了QSOX2、HTATIP2、UBE3C、MGST3、NSF、HECTD1、SAE1和ATG3参与下调UBL3与α-syn之间的相互作用。值得注意的是，沉默MGST3的影响最为显著。免疫细胞化学染色证实了MGST3沉默对细胞中UBL3与α-syn共定位的影响。MGST3是抗氧化系统的一部分，沉默MGST3被认为会导致氧化应激。我们用过氧化氢诱导氧化应激，观察其对UBL3-α-syn相互作用的影响，结果表明800µM的过氧化氢下调了这种相互作用。总之，沉默MGST3可下调UBL3与α-syn之间的相互作用。
