Abstract:
In recent years, the role of liquid-liquid phase separation (LLPS) and intrinsically disordered proteins (IDPs) in cellular molecular processes has received increasing attention from researchers. One such intrinsically disordered protein is TSPYL5, considered both as a marker and a potential therapeutic target for various oncological diseases. However, the role of TSPYL5 in intracellular processes remains unknown, and there is no clarity even in its intracellular localization. In this study, we characterized the intracellular localization and exchange dynamics with intracellular contents of TSPYL5 and its parts, utilizing TSPYL5 fusion proteins with EGFP. Our findings reveal that TSPYL5 can be localized in both the cytoplasm and nucleoplasm, including the nucleolus. The nuclear (nucleolar) localization of TSPYL5 is mediated by the nuclear/nucleolar localization sequences (NLS/NoLS) identified in the N-terminal intrinsically disordered region (4-27 aa), while its cytoplasmic localization is regulated by the ordered NAP-like domain (198-382 aa). Furthermore, our results underscore the significant role of the TSPYL5 N-terminal disordered region (1-198 aa) in the exchange dynamics with the nucleoplasm and its potential ability for phase separation. Bioinformatics analysis of the TSPYL5 interactome indicates its potential function as a histone and ribosomal protein chaperone. Taken together, these findings suggest a significant contribution of liquid-liquid phase separation to the processes involving TSPYL5, providing new insights into the role of this protein in the cell\'s molecular life.
摘要:
近年来,液-液相分离(LLPS)和内在无序蛋白(IDPs)在细胞分子过程中的作用越来越受到研究者的关注。一种这样的内在无序蛋白是TSPYL5,其被认为是各种肿瘤疾病的标志物和潜在治疗靶标。然而,TSPYL5在细胞内过程中的作用仍然未知,甚至在细胞内定位上也不清楚。在这项研究中,我们用TSPYL5及其部分的细胞内含量表征了细胞内定位和交换动力学,利用TSPYL5与EGFP的融合蛋白。我们的发现表明,TSPYL5可以定位在细胞质和核质中,包括核仁.TSPYL5的核(核仁)定位由在N末端固有无序区域(4-27aa)中鉴定的核/核仁定位序列(NLS/NoLS)介导,而其细胞质定位受有序NAP样结构域(198-382aa)调节。此外,我们的结果强调了TSPYL5N端无序区(1-198aa)在与核质的交换动力学中的重要作用及其潜在的相分离能力。TSPYL5相互作用组的生物信息学分析表明其作为组蛋白和核糖体蛋白伴侣的潜在功能。一起来看,这些发现表明液-液相分离对涉及TSPYL5的过程有重要贡献,为该蛋白在细胞分子生命中的作用提供了新的见解。