白假丝酵母Vps74蛋白的鉴定及其功能

Characterization of Vps74 in Candida albicans

摘要:

【背景】Vps74/GOLPH3是参与高尔基体蛋白糖基化修饰的关键蛋白，并且是重要的磷酸磷脂酰肌醇效应因子，在胞内参与多种信号通路。【目的】鉴定白假丝酵母Vps74蛋白，并探索其在该病原菌压力应答、蛋白分泌、形态发生及致病过程中的功能。【方法】采用在线序列比对方法，初步鉴定白假丝酵母Vps74蛋白；采用两步PCR介导的同源重组方法，构建白假丝酵母vps74基因缺失菌株vps74Δ/Δ及回补菌株VPS74c；采用反向遗传学方法，探究Vps74在白假丝酵母的压力应答、蛋白分泌、形态发生及致病过程中的功能。【结果】白假丝酵母中存在典型的Vps74/GOLPH3同源蛋白，Vps74参与蛋白糖基化修饰过程，vps74基因缺失导致白假丝酵母蛋白分泌能力、形态发生能力、黏附能力以及侵染宿主能力的显著降低。【结论】Vps74通过影响蛋白分泌、形态发生、黏附、嵌入式生长等过程，在白假丝酵母致病过程中发挥重要作用。

Abstract:

[Background] Vps74/GOLPH3 is a key protein involved in the glycosylation of the Golgi protein, and an important phosphatidylinositol-4-kinase effector and phosphatidylinositol-4- phosphate sensor participating in a variety of intracellular signaling pathways. [Objective] To characterize the Vps74/GOLPH3 homologue in Candida albicans and investigate the role of Cavps74 in stress response, secretion, morphogenesis and pathogenicity. [Methods] Sequence alignment and analysis were performed by the relevant biosoftware. The gene deletion strain vps74?/? and reconstructed strain VPS74c were constructed by PCR-mediated gene disruption. With reverse genetic analysis, we further investigated the function of Vps74 in protein glycosylation, stress response, secretion, morphogenesis and pathogenicity. [Results] There is a typical homologue of Vps74/GOLPH3 in C. albicans which also plays important role in glycosylation process. Deletion of vps74 resulted in various effects, including decreased secretion ability, morphogenesis, adhesion and loss of pathogenicity. [Conclusion] Vps74 plays a key role in protein secretion, morphogenesis, adhesion and pathogenicity in C. albicans.