Estimating the expression levels of genes controlling biofilm formation and evaluating the effects of different conditions on biofilm formation and secreted aspartic proteinase activity in Candida albicans and Saccharomyces cerevisiae: a comparative study

IF 2.5 Q2 MULTIDISCIPLINARY SCIENCES
Shaimaa S. Sobieh, Rowida G. Elshazly, Sahar A. Tawab, Sanaa S. Zaki
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引用次数: 0

Abstract

Background

Characterization of yeast virulence genes is an important tool for identifying the molecular pathways involved in switching yeast virulence. Biofilm formation (BF) and secreted aspartic proteinase (SAP) activity are essential virulence factors that contribute to yeast pathogenicity.

Results

Four Candida albicans and two Saccharomyces cerevisiae strains were tested for BF and SAP activity under optimum conditions, and the expression levels of several genes controlling BF were quantified under the optimal conditions. Biofilm formation was assessed by the microplate method at different pH values, incubation times and culture media. Similarly, SAP activity was assessed at different pH values and incubation periods. The expression levels of nine genes were determined via qRT-PCR technique. All tests were carried out in triplicate, and the values presented as the means ± standard deviations and were analysed with the SPSS programme. Only C. albicans (1), C. albicans (2) and S. cerevisiae 43 formed biofilms. The optimal BF was obtained after culture in sabouraud dextrose broth with 8% glucose at pH 7.5, 4 and 6, respectively, for 48h. Candida albicans biofilm production was more significant than that of S. cerevisiae 43. Moreover, the SAP activity was estimated under the optimum conditions. All yeasts showed optimal SAP activity at pH 4, but astonishingly the SAP activity of S. cerevisiae 44 was higher than that of C. albicans. The expression levels of EFG1 and ZAP1 (transcription factors); ALS3, HWP1and YWP1 (adhesion genes); SAP1 and SAP4 (aspartic proteinase) in C. albicans (1); and FLO11 (adhesion gene) and YPS3 (aspartic proteinase) in S. cerevisiae 43 were quantified during biofilm development at different time intervals. The expression levels of EFG1, ALS3, YWP1, SAP1, SAP4, FLO11 and YPS3 were upregulated at 8 h, while that of ZAP1 was upregulated at 48 h. Only HWP1 was downregulated.

Conclusions

The findings of the present study may provide information for overcoming yeast BF and pathogenicity by regulating specific genes at specific times. Additionally, this study revealed the virulence of the commensal S. cerevisiae, which may take the pathogenicity direction as C. albicans.

估算控制白念珠菌和酿酒酵母生物膜形成的基因表达水平以及评估不同条件对白念珠菌生物膜形成和分泌天冬氨酸蛋白酶活性的影响:一项比较研究
背景酵母毒力基因的表征是确定参与酵母毒力转换的分子途径的重要工具。结果对四株白念珠菌和两株酿酒酵母在最佳条件下的生物膜形成(BF)和分泌天冬氨酸蛋白酶(SAP)活性进行了检测,并对控制BF的几个基因在最佳条件下的表达水平进行了量化。在不同的 pH 值、培养时间和培养基条件下,采用微孔板法对生物膜的形成进行了评估。同样,在不同的 pH 值和培养时间下也对 SAP 活性进行了评估。通过 qRT-PCR 技术测定了九种基因的表达水平。所有测试均一式三份,数值以平均值±标准偏差表示,并使用 SPSS 程序进行分析。只有白僵菌(1)、白僵菌(2)和 S. cerevisiae 43 形成了生物膜。在 pH 值分别为 7.5、4 和 6 的含 8%葡萄糖的沙保葡萄糖肉汤中培养 48 小时后,获得了最佳生物膜。白色念珠菌生物膜的产生比 S. cerevisiae 43 更为显著。此外,还对最佳条件下的 SAP 活性进行了评估。所有酵母菌都在 pH 值为 4 时表现出最佳的 SAP 活性,但令人惊讶的是,酿酒酵母菌 44 的 SAP 活性高于白念珠菌。在生物膜形成过程中,对不同时间间隔内白僵菌(1)中的 EFG1 和 ZAP1(转录因子);ALS3、HWP1 和 YWP1(粘附基因);SAP1 和 SAP4(天冬氨酸蛋白酶);以及 S. cerevisiae 43 中的 FLO11(粘附基因)和 YPS3(天冬氨酸蛋白酶)的表达水平进行了量化。结论本研究的结果可为通过在特定时间调控特定基因来克服酵母生物膜和致病性提供信息。此外,本研究还揭示了共生酵母菌 S. cerevisiae 的致病性,它可能会像白僵菌一样走上致病的道路。
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来源期刊
CiteScore
2.60
自引率
0.00%
发文量
0
期刊介绍: Beni-Suef University Journal of Basic and Applied Sciences (BJBAS) is a peer-reviewed, open-access journal. This journal welcomes submissions of original research, literature reviews, and editorials in its respected fields of fundamental science, applied science (with a particular focus on the fields of applied nanotechnology and biotechnology), medical sciences, pharmaceutical sciences, and engineering. The multidisciplinary aspects of the journal encourage global collaboration between researchers in multiple fields and provide cross-disciplinary dissemination of findings.
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