Kundu Gautam, Ghasemi Maryam, Yim Seungbin, Rohil Ayanna, Xin Cuiyan, Ren Leo, Srivastava Shraddha, Akinfolarin Akinwande, Kumar Subodh, Srivastava Gyan P, Sabbisetti Venkata S, Murugaiyan Gopal, Ajay Amrendra K
Division of Renal Medicine, Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.
US Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA.
Biomedicines. 2024 Sep 14;12(9):2102. doi: 10.3390/biomedicines12092102.
Signal transducer and activator of transcription 3 (STAT3) is a member of the cytoplasmic inducible transcription factors and plays an important role in mediating signals from cytokines, chemokines, and growth factors. We and others have found that STAT3 directly regulates pro-fibrotic signaling in the kidney. The STAT3 protein-protein interaction plays an important role in activating its transcriptional activity. It is necessary to identify these interactions to investigate their function in kidney disease. Here, we investigated the protein-protein interaction among three species to find crucial interactions that can be targeted to alleviate kidney disease.
In this study, we examined common protein-protein interactions leading to the activation or downregulation of STAT3 among three different species: humans (), mice (), and rabbits (). Further, we chose to investigate the P300 and STAT3 interaction and performed studies of the activation of STAT3 using IL-6 and inhibition of the P300 by its specific inhibitor A-485 in pericytes. Next, we performed immunoprecipitation to confirm whether A-485 inhibits the binding of P300 to STAT3.
Using the STRING application from ExPASy, we found that six proteins, including PIAS3, JAK1, JAK2, EGFR, SRC, and EP300, showed highly confident interactions with STAT3 in humans, mice, and rabbits. We also found that IL-6 treatment increased the acetylation of STAT3 and increased histone 3 lysine acetylation (H3K27ac). Furthermore, we found that the disruption of STAT3 and P300 interaction by the P300 inhibitor A-485 decreased STAT3 acetylation and H3K27ac. Finally, we confirmed that the P300 inhibitor A-485 inhibited the binding of STAT3 with P300, which inhibited its transcriptional activity by reducing the expression of ().
Targeting the P300 protein interaction with STAT3 may alleviate STAT3-mediated fibrotic signaling in humans and other species.
信号转导与转录激活因子3（STAT3）是细胞质诱导型转录因子成员，在介导细胞因子、趋化因子和生长因子的信号传导中起重要作用。我们及其他人发现STAT3直接调节肾脏中的促纤维化信号传导。STAT3蛋白-蛋白相互作用在激活其转录活性中起重要作用。识别这些相互作用对于研究它们在肾脏疾病中的功能很有必要。在此，我们研究了三种物种之间的蛋白-蛋白相互作用，以找到可作为靶点来减轻肾脏疾病的关键相互作用。
在本研究中，我们检测了导致三种不同物种（人类、小鼠和兔子）中STAT3激活或下调的常见蛋白-蛋白相互作用。此外，我们选择研究P300与STAT3的相互作用，并在周细胞中使用白细胞介素-6进行STAT3激活研究以及用其特异性抑制剂A-485抑制P300。接下来，我们进行免疫沉淀以确认A-485是否抑制P300与STAT3的结合。
使用ExPASy的STRING应用程序，我们发现包括PIAS3、JAK1、JAK2、EGFR、SRC和EP300在内的六种蛋白质在人类、小鼠和兔子中与STAT3表现出高度可靠的相互作用。我们还发现白细胞介素-6处理增加了STAT3的乙酰化并增加了组蛋白3赖氨酸乙酰化（H3K27ac）。此外，我们发现P300抑制剂A-485破坏STAT3与P300的相互作用会降低STAT3乙酰化和H3K27ac。最后，我们证实P300抑制剂A-485抑制了STAT3与P300的结合，这通过降低（）的表达抑制了其转录活性。
靶向P300与STAT3的蛋白相互作用可能减轻人类和其他物种中STAT3介导的纤维化信号传导。
