Bernardo Stella M, Rane Hallie S, Chavez-Dozal Alba, Lee Samuel A
Section of Infectious Diseases, New Mexico Veterans Healthcare System, Albuquerque, NM, USA; University of New Mexico Health Science Center, Albuquerque, NM, USA.
FEMS Yeast Res. 2014 Aug;14(5):762-75. doi: 10.1111/1567-1364.12165. Epub 2014 Jun 9.
To study the role of late secretion in Candida albicans pathogenesis, we created conditional mutant C. albicans strains in which the t-SNARE-encoding genes SSO2 or SEC9 were placed under the control of a tetracycline-regulated promoter. In repressing conditions, C. albicans tetR-SSO2 and tetR-SEC9 mutant strains were defective in cytokinesis and secretion of aspartyl proteases and lipases. The mutant strains also exhibited a defect in filamentation compared with controls, and thus, we followed the fate of the C. albicans Spitzenkörper, an assembly of secretory vesicles thought to act as a vesicle supply center for the growing hyphae. In the absence of Ca Sso2p, the Spitzenkörper dissipated within 5 h and thin-section electron microscopy revealed an accumulation of secretory vesicles. Moreover, the hyphal tip developed into a globular yeast-like structure rather than maintaining a typical narrow hyphae. These studies indicate that late secretory t-SNARE proteins in C. albicans are required for fundamental cellular processes and contribute to virulence-related attributes of C. albicans pathogenesis. Moreover, these results provide direct evidence for a key role of SNARE proteins in vesicle-mediated polarized hyphal growth of C. albicans.
为了研究晚期分泌在白色念珠菌致病机制中的作用,我们构建了条件突变的白色念珠菌菌株,其中编码t-SNARE的基因SSO2或SEC9置于四环素调控启动子的控制之下。在抑制条件下,白色念珠菌tetR-SSO2和tetR-SEC9突变菌株在胞质分裂以及天冬氨酸蛋白酶和脂肪酶的分泌方面存在缺陷。与对照相比,突变菌株在菌丝形成方面也表现出缺陷,因此,我们追踪了白色念珠菌Spitzenkörper的命运,Spitzenkörper是分泌囊泡的集合体,被认为是生长中菌丝的囊泡供应中心。在缺乏Ca Sso2p的情况下,Spitzenkörper在5小时内消散,超薄切片电子显微镜显示分泌囊泡积累。此外,菌丝尖端发育成球状酵母样结构,而不是维持典型的狭窄菌丝。这些研究表明,白色念珠菌中的晚期分泌t-SNARE蛋白是基本细胞过程所必需的,并有助于白色念珠菌致病机制中与毒力相关的特性。此外,这些结果为SNARE蛋白在白色念珠菌囊泡介导的极性菌丝生长中的关键作用提供了直接证据。