Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo 04023-062, Brazil.
Laboratório Especial de Ciclo Celular-Center of Toxins, Immune Response and Cell Signaling-Center (CeTICS), Butantan Institute, São Paulo 05503-900, Brazil.
Cells. 2019 Jul 23;8(7):765. doi: 10.3390/cells8070765.
and cause human paracoccidioidomycosis. We have previously characterized the <200-nt RNA sub-populations contained in fungal extracellular vesicles (EVs) from Pb18 and other pathogenic fungi. We have presently used the RNA-seq strategy to compare the <200- and >200-nt RNA fractions contained in EVs isolated from culture supernatants of Pb18, Pb3, and Pb01. Shared mRNA sequences were related to protein modification, translation, and DNA metabolism/biogenesis, while those related to transport and oxidation-reduction were exclusive to Pb01. The presence of functional full-length mRNAs was validated by in vitro translation. Among small non-coding (nc)RNA, 15 were common to all samples; small nucleolar (sno)RNAs were enriched in EVs, whereas for there were similar proportions of snoRNA, rRNA, and tRNA. Putative exonic sRNAs were highly abundant in Pb18 EVs. We also found sRNA sequences bearing incomplete microRNA structures mapping to exons. RNA-seq data suggest that extracellular fractions containing Pb18 EVs can modulate the transcriptome of murine monocyte-derived dendritic cells in a transwell system. Considering that sRNA classes are involved in transcription/translation modulation, our general results may indicate that differences in virulence among fungal isolates can be related to their distinct EV-RNA content.
并导致人类副球孢子菌病。我们之前已经对 Pb18 和其他致病真菌的细胞外囊泡 (EV) 中包含的<200nt RNA 亚群进行了特征描述。目前，我们使用 RNA-seq 策略比较了从 Pb18、Pb3 和 Pb01 的培养上清液中分离的 EV 中包含的<200nt 和>200nt RNA 分数。共享的 mRNA 序列与蛋白质修饰、翻译和 DNA 代谢/生物发生有关，而与运输和氧化还原相关的序列则仅存在于 Pb01 中。通过体外翻译验证了功能性全长 mRNA 的存在。在小非编码 (nc)RNA 中，有 15 种在所有样本中都存在；小核仁 (sno)RNA 在 EVs 中富集，而在 Pb01 中 snoRNA、rRNA 和 tRNA 的比例相似。外显子 sRNA 在 Pb18 EVs 中高度丰富。我们还发现了具有不完整 miRNA 结构的 sRNA 序列，这些序列映射到外显子上。RNA-seq 数据表明，含有 Pb18 EV 的细胞外成分可以在 Transwell 系统中调节鼠单核细胞来源的树突状细胞的转录组。考虑到 sRNA 类参与转录/翻译调节，我们的总体结果可能表明真菌分离株之间毒力的差异可能与其独特的 EV-RNA 含量有关。
