Department of Pathogenic Biology and Immunology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, No.76 Yanta West Road, Xi'an, 710061, China.
Clinical Laboratory Center, Xi'an People's Hospital, Xi'an Fourth Hospital, Affiliated People's Hospital of Northwest University, Xi'an, 710004, China.
BMC Genomics. 2024 Aug 28;25(1):811. doi: 10.1186/s12864-024-10715-2.
Dermatophagoides farinae proteins (DFPs) are abundantly expressed in D. farinae; however, their functions remain unknown. Our previous transcriptome sequencing analyses revealed that the basal expression of DFP1 and DFP2 in D. farinae was high and, more importantly, upregulated under temperature stress. Therefore, DFPs were speculated to exert a temperature stress response function.
Real-time quantitative polymerase chain reaction detection revealed that both DFP1 and DFP2 were significantly upregulated under temperature stress. Particularly, DFP1 was upregulated under cold stress. Electrophoresis of D. farinae total proteins revealed an increased abundance of DFP1 and DFP2 (40-55 kDa bands) under temperature stress, which was corroborated by the mass spectrometry results. After silencing DFP1 and DFP2 further, temperature stress led to decreases in gene expression and survival rates. Moreover, DFP1 was identified as the upstream regulator of DFP2.
This study highlights the temperature stress response functions of DFP1 and DFP2 at the mRNA and protein levels. These results provide important insights for applying DFP1 and DFP2 as potential target genes for the molecular prevention and control of D. farinae to prevent allergic diseases. The newly established methods provide methodological guidance for the study of genes with unknown functions in mites.
粉尘螨蛋白(DFP)在粉尘螨中大量表达,但它们的功能尚不清楚。我们之前的转录组测序分析表明,DFP1 和 DFP2 在粉尘螨中的基础表达水平较高,更重要的是,在温度胁迫下上调。因此,推测 DFP 具有温度应激反应功能。
实时定量聚合酶链反应检测显示,DFP1 和 DFP2 在温度胁迫下均显著上调。特别是 DFP1 在冷胁迫下上调。粉尘螨总蛋白电泳显示,DFP1 和 DFP2(40-55 kDa 带)在温度胁迫下丰度增加,质谱结果也得到了证实。进一步沉默 DFP1 和 DFP2 后,温度应激导致基因表达和存活率下降。此外,DFP1 被鉴定为 DFP2 的上游调控因子。
本研究在 mRNA 和蛋白质水平上强调了 DFP1 和 DFP2 的温度应激反应功能。这些结果为应用 DFP1 和 DFP2 作为粉尘螨分子预防和控制的潜在靶基因,以预防过敏疾病提供了重要的见解。新建立的方法为研究螨虫中功能未知的基因提供了方法学指导。