Ueda Ryota, Hashimoto Rina, Fujii Yuki, Menezes José C J M D S, Takahashi Hirotaka, Takeda Hiroyuki, Sawasaki Tatsuya, Motokawa Tomonori, Tokunaga Kenzo, Fujita Hideaki
Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo 859-3298, Japan.
Esteem Industries Pvt Ltd., Bicholim 403529, Goa, India.
Membranes (Basel). 2024 Feb 1;14(2):43. doi: 10.3390/membranes14020043.
Lysosomal degradation of tyrosinase, a pivotal enzyme in melanin synthesis, negatively impacts melanogenesis in melanocytes. Nevertheless, the precise molecular mechanisms by which lysosomes target tyrosinase have remained elusive. Here, we identify RING (Really Interesting New Gene) finger protein 152 (RNF152) as a membrane-associated ubiquitin ligase specifically targeting tyrosinase for the first time, utilizing AlphaScreen technology. We observed that modulating RNF152 levels in B16 cells, either via overexpression or siRNA knockdown, resulted in decreased or increased levels of both tyrosinase and melanin, respectively. Notably, RNF152 and tyrosinase co-localized at the trans-Golgi network (TGN). However, upon treatment with lysosomal inhibitors, both proteins appeared in the lysosomes, indicating that tyrosinase undergoes RNF152-mediated lysosomal degradation. Through ubiquitination assays, we found the indispensable roles of both the RING and transmembrane (TM) domains of RNF152 in facilitating tyrosinase ubiquitination. In summary, our findings underscore RNF152 as a tyrosinase-specific ubiquitin ligase essential for regulating melanogenesis in melanocytes.
酪氨酸酶是黑色素合成中的关键酶,其溶酶体降解对黑素细胞中的黑色素生成产生负面影响。然而,溶酶体靶向酪氨酸酶的确切分子机制仍不清楚。在这里,我们首次利用AlphaScreen技术鉴定出RING(真有趣新基因)指蛋白152(RNF152)是一种特异性靶向酪氨酸酶的膜相关泛素连接酶。我们观察到,通过过表达或siRNA敲低来调节B16细胞中的RNF152水平,分别导致酪氨酸酶和黑色素水平的降低或升高。值得注意的是,RNF152和酪氨酸酶共定位于反式高尔基体网络(TGN)。然而,在用溶酶体抑制剂处理后,这两种蛋白都出现在溶酶体中,表明酪氨酸酶经历了RNF152介导的溶酶体降解。通过泛素化分析,我们发现RNF152的RING结构域和跨膜(TM)结构域在促进酪氨酸酶泛素化方面都起着不可或缺的作用。总之,我们的研究结果强调了RNF152作为一种酪氨酸酶特异性泛素连接酶,对调节黑素细胞中的黑色素生成至关重要。