División de Biología Molecular, IPICYT, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José, San Luis Potosí, San Luis Potosí, Mexico.
Mycopathologia. 2013 Apr;175(3-4):207-19. doi: 10.1007/s11046-013-9627-2. Epub 2013 Feb 8.
The opportunistic fungal pathogen Candida glabrata adheres tightly to epithelial cells in culture, mainly through the adhesin Epa1. EPA1 is the founding member of a family of up to 23 putative adhesin-encoding genes present in the C. glabrata genome. The majority of the EPA genes are localized close to the telomeres, where they are repressed by subtelomeric silencing that depends on the Sir, Ku, Rif1, and Rap1 proteins. EPA6 and EPA7 also encode functional adhesins that are repressed in vitro. EPA1 expression in vitro is tightly controlled both positively and negatively, and in addition, presents high cell-to-cell heterogeneity, which depends on Sir-mediated silencing. In this work, we characterized the ability to adhere to HeLa epithelial cells and the expression of several EPA genes in a collection of 79 C. glabrata clinical isolates from several hospitals in Mexico. We found 11 isolates that showed increased adherence to mammalian cells compared with our reference strain under conditions where EPA1 is not expressed. The majority of these isolates displayed over-expression of EPA1 and EPA6 or EPA7, but did not show increased biofilm formation. Sequencing of the SIR3 gene of several hyper-adherent isolates revealed that all of them contain several polymorphisms with respect to the reference strain. Interestingly, two isolates have polymorphisms in positions flanked by clusters of amino acids required for silencing in the Saccharomyces cerevisiae Sir3 protein. Our data show that there is a large variability in adhesin expression and adherence to epithelial cells among different C. glabrata clinical isolates.
机会性真菌病原体 Candida glabrata 在培养物中紧密附着于上皮细胞,主要通过黏附素 Epa1 实现。EPA1 是一个包含多达 23 个假定黏附素编码基因的家族的创始成员,这些基因存在于 C. glabrata 基因组中。大多数 EPA 基因位于端粒附近,在端粒处受到端粒沉默的抑制,这种抑制依赖于 Sir、Ku、Rif1 和 Rap1 蛋白。EPA6 和 EPA7 也编码功能性黏附素,在体外受到抑制。EPA1 在体外的表达受到严格的正调控和负调控,此外,还表现出高度的细胞间异质性,这依赖于 Sir 介导的沉默。在这项工作中,我们对来自墨西哥几家医院的 79 株 C. glabrata 临床分离株的粘附上皮细胞的能力和几个 EPA 基因的表达进行了表征。我们发现,在不表达 EPA1 的情况下,与我们的参考菌株相比,有 11 株分离株在粘附哺乳动物细胞方面表现出增强的能力。这些分离株中的大多数表现出 EPA1 和 EPA6 或 EPA7 的过度表达,但没有表现出增加的生物膜形成。对几个高粘附分离株的 SIR3 基因进行测序发现,它们都含有相对于参考菌株的多个多态性。有趣的是,两个分离株在沉默所必需的氨基酸簇侧翼的位置存在多态性,在酿酒酵母 Sir3 蛋白中。我们的数据表明,不同 C. glabrata 临床分离株之间的黏附素表达和对上皮细胞的粘附存在很大的可变性。