UBE2O ubiquitinates PTRF/CAVIN1 and inhibits the secretion of exosome-related PTRF/CAVIN1.
UBE2O ubiquitinates PTRF/CAVIN1 and inhibits the secretion of exosome-related PTRF/CAVIN1.
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UBE2O 泛素化 PTRF/CAVIN1 并抑制外泌体相关 PTRF/CAVIN1 的分泌
DOI：
10.1186/s12964-022-00996-z
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发表时间：
2022-11-28
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外泌体是细胞释放的小囊泡，在细胞间通讯中具有关键作用。外泌体起源于细胞膜内陷，并在多囊泡体（MVBs）与细胞膜融合后被释放。已知聚合酶I和转录释放因子（PTRF，也称为小窝蛋白相关蛋白 - 1，CAVIN1）在小窝形成和外泌体分泌中起重要作用。并且外泌体中的PTRF已被确定为多种恶性肿瘤（如神经胶质瘤和肾细胞癌）的潜在生物标志物。然而，如何调节外泌体相关PTRF分泌的机制仍然未知。
我们进行了外源性和内源性免疫沉淀试验，以研究泛素结合酶E2O（UBE2O）与PTRF之间的相互作用。我们通过泛素化试验确定了UBE2O使PTRF泛素化。然后，通过超速离心分离外泌体，并通过透射电子显微镜、蛋白质印迹和纳米颗粒跟踪分析进行鉴定。通过蛋白质印迹分析UBE2O对外泌体相关PTRF分泌的影响，并通过外泌体标志物和外泌体总蛋白含量评估UBE2O对外泌体分泌的影响。
在此，我们表明UBE2O直接与PTRF相互作用并使PTRF泛素化。在功能上，我们发现UBE2O通过减少小窝形成抑制了PTRF对外泌体分泌的影响。重要的是，UBE2O减少了外泌体分泌，从而通过外泌体下调了PTRF的分泌。我们的研究还确定了血清剥夺蛋白反应（SDPR，也称为小窝蛋白相关蛋白 - 2，CAVIN2）与UBE2O和PTRF都相互作用。此外，我们发现SDPR促进外泌体中PTRF的表达。有趣的是，即使存在SDPR，UBE2O仍然抑制外泌体相关PTRF的分泌。
我们的研究表明，UBE2O通过使PTRF泛素化而下调外泌体释放并控制外泌体相关PTRF的分泌。由于外泌体在恶性肿瘤生长中起重要作用，并且外泌体中包含的PTRF是几种恶性肿瘤的生物标志物，增加细胞中UBE2O的表达有可能被开发为一种新的癌症治疗方法。
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在线版本包含补充材料，可在10.1186/s12964 - 022 - 00996 - z获取。
Exosomes are small vesicles released by cells, which have crucial functions in intercellular communication. Exosomes originated from cell membrane invagination and are released followed by multivesicular bodies (MVBs) fused with the cell membrane. It is known that Polymerase I and Transcript Release Factor (PTRF, also known as Caveolin-associated Protein-1, CAVIN1) plays an important role in caveolae formation and exosome secretion. And PTRF in exosomes has been identified as a potential biomarker in multiple malignancies such as glioma and renal cell carcinoma. However, the mechanisms of how to regulate the secretion of exosome-related PTRF remain unknown. We performed exogenous and endogenous immunoprecipitation assays to investigate the interaction between ubiquitin-conjugating enzyme E2O (UBE2O) and PTRF. We identified UBE2O ubiquitinated PTRF using ubiquitination assays. Then, exosomes were isolated by ultracentrifugation and identified by transmission electronic microscopy, western blot and nanoparticle tracking analysis. The effect of UBE2O on the secretion of exosome-related PTRF was analyzed by western blot, and the effect of UBE2O on exosome secretion was evaluated by exosome markers and the total protein content of exosomes. Here, we showed that UBE2O interacts with PTRF directly and ubiquitinates PTRF. Functionally, we found that UBE2O inhibited the effects of PTRF on exosome secretion via decreasing caveolae formation. Importantly, UBE2O decreased exosome secretion, resulting in downregulating PTRF secretion via exosomes. Our study also identified Serum Deprivation Protein Response (SDPR, also known as Caveolin-associated Protein-2, CAVIN2) interacted with both UBE2O and PTRF. Furthermore, we found that SDPR promotes PTRF expression in exosomes. Interestingly, even in the presence of SDPR, UBE2O still inhibited the secretion of exosome-related PTRF. Our study demonstrated that UBE2O downregulated exosome release and controlled the secretion of exosome-related PTRF through ubiquitinating PTRF. Since exosomes play an important role in malignant tumor growth and PTRF included in exosomes is a biomarker for several malignant tumors, increasing UBE2O expression in cells has the potential to be developed as a novel approach for cancer treatment. Video Abstract The online version contains supplementary material available at 10.1186/s12964-022-00996-z.
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