Yang Longbing, Tian Zhuqing, Zhou Luoxiong, Sun Chaoqin, Huang Mingjiao, Tian Chunren, Peng Jian, Guo Guo
The Key and Characteristic Laboratory of Modern Pathogen Biology, School of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou, China.
Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, Guizhou, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Jan 25;39(1):304-317. doi: 10.13345/j.cjb.220499.
is one of the major causes of invasive fungal infections and a serious opportunistic pathogen in immunocompromised individuals. The antimicrobial peptide AMP-17 has prominent anti- activity, and proteomic analysis revealed significant differences in the expression of cell wall () and oxidative stress () genes upon the action of AMP-17 on . , suggesting that AMP-17 may exert anti-. effects by affecting the expression of and genes. To further investigate whether and genes were the targets of AMP-17, . and mutants were constructed using the clustered regulatory interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system. Phenotypic observations revealed that deletion of two genes had no significant effect on . growth and biofilm formation, whereas gene deletion affected stress response and mycelium formation of . . Drug sensitivity assay showed that the MIC values of AMP-17 against and mutants increased from 8 μg/mL (for the wild type . SC5314) to 16 μg/mL, while the MIC values against : : revertants decreased to the level of the wild type SC5314. In addition, the ability of AMP-17 to inhibit biofilm formation of both deletion strains was significantly reduced compared to that of wild type SC5314, indicating that the susceptibility of the deletion mutants to AMP-17 was reduced in both the yeast state and during biofilm formation. These results suggest that and genes are likely two of the potential targets for AMP-17 to exert anti. effects, which may facilitate further exploration of the antibacterial mechanism of novel peptide antifungal drugs.
是侵袭性真菌感染的主要原因之一，也是免疫功能低下个体中的一种严重机会性病原体。抗菌肽AMP - 17具有显著的抗活性，蛋白质组学分析显示，在AMP - 17作用于 后，细胞壁（）和氧化应激（）相关基因的表达存在显著差异。 ，这表明AMP - 17可能通过影响 和 基因的表达发挥抗 作用。为了进一步研究 和 基因是否是AMP - 17的作用靶点，使用成簇规律间隔短回文重复序列相关蛋白9（CRISPR/Cas9）系统构建了 和 突变体。表型观察表明，两个基因的缺失对 的生长和生物膜形成没有显著影响，而 基因的缺失影响了 的应激反应和菌丝体形成。药敏试验表明，AMP - 17对 和 突变体的最低抑菌浓度（MIC）值从8μg/mL（野生型 SC5314）增加到16μg/mL，而对 : : 回复菌株的MIC值则降至野生型SC5314的水平。此外，与野生型SC5314相比，AMP - 17抑制两种缺失菌株生物膜形成的能力显著降低，这表明缺失突变体在酵母状态和生物膜形成过程中对AMP - 17的敏感性均降低。这些结果表明， 和 基因可能是AMP - 17发挥抗 作用的两个潜在靶点，这可能有助于进一步探索新型肽类抗真菌药物的抗菌机制。
