脑炎微孢子虫与宿主线粒体VDAC蛋白的互作机制研究

Microsporidia are unicellular, obligate intracellular fungal parasites. The majority species of microsporidia that can infect humans are Encephalitozoonidae. As the microsporidia lacking functional mitochondria and almost all of the genes for ATP generation, the invasion and reproduction of microsporidia were closely associated with host mitochondria. It was reported that the voltage-dependent anion-selective channel protein (VDAC) on the outer membrane of host cell mitochondria participates in the process of the microsporidia and host mitochondria interaction. However, the underlying molecular mechanisms remain unclear. Our preliminary studies found that the downregulation of the VDAC expression can significantly inhibit the host mitochondria and parasite association, and the microsporidia sporoplasm surface protein 1 (SSP1) was proved to be one of the interacting proteins of VDAC. Thus, we hypothesize that VDAC may play a critical role in the interaction of microsporidia and host mitochondria. We aim to: (1) Search for the VDAC interacting target proteins on microsporidia based on the TurboID proximity biotinylation technology; (2) Prove the interaction of VDAC and its target proteins by Yeast Two-hybrid and fluorescence colocalization technologies; (3) Illuminate the function of VDAC and its interacting proteins during the process of microsporidia infection and cell development. The successful completion of this project will improve the current understanding of the mechanisms of microsporidia and host mitochondria interaction, and the information gained from this study will provide new therapeutic targets for the control and prevention of microsporidiosis.

微孢子虫是一种专性细胞内寄生的单细胞病原真菌，由于该病原缺少线粒体以及能量合成相关基因，因此微孢子虫的感染和增殖与宿主线粒体密切相关，研究表明宿主线粒体外膜电压依赖性阴离子通道蛋白（VDAC）参与宿主线粒体与微孢子虫的互作过程，但其分子机制尚不清楚。预实验发现下调宿主VDAC的表达能够明显抑制线粒体与病原的结合，且VDAC与微孢子虫孢原质表面蛋白SSP1存在互作。为此我们推测VDAC是宿主线粒体与微孢子虫互作过程中的关键因子。本项目拟（1）采用TurboID生物素标记法对与VDAC存在互作的微孢子虫蛋白进行筛选；（2）利用酵母双杂交实验以及荧光共定位实验等对VDAC及其互作蛋白进行验证；（3）通过抗体封闭实验等对VDAC及其互作蛋白在微孢子虫侵染和繁殖过程中的作用进行探讨。本项目将有助于进一步理解微孢子虫与宿主线粒体的互作机制，为微孢子虫病的预防和治疗提供新的治疗靶标。

内容获取失败，请点击重试