中华皮肤科杂志 ›› 2019, Vol. 52 ›› Issue (3): 176-181.doi: 10.3760/cma.j.issn.0412-4030.2019.03.006

• 论著 • 上一篇    下一篇

申克孢子丝菌酵母相形成过程中转录组表达谱变化及差异表达基因功能分析

韩昌旭1    侯彬彬2    晋亮3    张振颖1   

  1. 1香港大学深圳医院皮肤科 518000; 2大连医科大学附属第二医院皮肤科 116000; 3空军特色医学中心皮肤科,北京 100142
  • 收稿日期:2018-06-12 修回日期:2018-12-12 出版日期:2019-03-15 发布日期:2019-03-04
  • 通讯作者: 张振颖 E-mail:zhangzy@hku-szh.org
  • 基金资助:
    国家自然科学基金(81472891、81502737)

Changes in transcriptome expression profiles and functional analysis of differentially expressed genes during mycelium-to-yeast transformation of Sporothrix schenckii

Han Changxu1, Hou Binbin2, Jin Liang3, Zhang Zhenying1   

  1. 1Department of Dermatology, The University of Hong Kong⁃Shenzhen Hospital, Shenzhen 518000, China; 2Department of Dermatology, The Second Hospital of Dalian Medical University, Dalian 116000, China; 3Department of Dermatology, Air Force Medical Center, Beijing 100142, China
  • Received:2018-06-12 Revised:2018-12-12 Online:2019-03-15 Published:2019-03-04
  • Contact: Zhang Zhen-ying E-mail:zhangzy@hku-szh.org
  • Supported by:
    National Natural Science Foundation of China (81472891, 81502737)

摘要: 【摘要】 目的 比较并分析申克孢子丝菌菌丝相及早期酵母相转录组学的差异,明确酵母相形成过程中转录组表达谱的变化。方法 对25 ℃沙氏培养基培养96 h获得的菌丝相申克孢子丝菌及37 ℃脑心浸液培养基培养36 h获得的早期酵母相申克孢子丝菌的转录组进行测序分析,对筛选后的单基因簇进行功能注释,包括与多个数据库(NR、Swiss?Prot、KEGG、COG、KOG、GO、Pfam)的比对、编码区序列预测及基因在每个样品中的表达量分析。最后,对差异表达基因进行模式聚类、功能注释以及富集性分析。结果 共获得14.76 Gb有效数据,组装获得43 863条单基因簇,28 094条获得注释结果。与菌丝相比较,酵母相上调表达的基因10 969条,下调表达的基因199条。差异表达基因参与蛋白磷酸化、细胞内蛋白转运、细胞蛋白修饰、小三磷酸鸟苷酶介导的信号转导、囊泡介导转运、翻译、细胞内信号转导、微管形成、三磷酸腺苷合成偶联质子转运等过程。参与真菌形态形成的两个重要信号转导通路丝裂原活化蛋白激酶和双组分信号通路中的16条基因及参与几丁质合成代谢的16条基因均被证实在早期酵母相上调表达。结论 与菌丝相相比,申克孢子丝菌酵母相基因表达谱发生显著变化,差异表达的基因涉及众多功能,提示孢子丝菌相变受多基因网络系统的调控。

关键词: 孢子丝菌属, 菌丝, 转录组, 基因表达谱, 差异表达基因分析, 酵母相

Abstract: 【Abstract】 Objective To compare and analyze the differences in the transcriptomics between mycelium and early yeast phases of Sporothrix schenckii (S. schenckii), and to realize the changes in transcriptome expression profiles during mycelium-to-yeast transformation. Methods A standard strain of S. schenckii (ATCC 10268) was subjected to 96-hour culture with Sabouraud medium at 25 ℃ or 36-hour culture with brain-heart infusion medium at 37 ℃ to obtain the mycelium and yeast form of S. schenckii, and then, their transcriptomes were sequenced. Functional annotation was performed for screened unigenes by comparison using several databases (such as NR, Swiss-Prot, KEGG, COG, KOG, GO and Pfam), coding sequence prediction, and gene expression analysis in each sample. Finally, the differentially expressed genes were subjected to pattern clustering, functional annotation and enrichment analysis. Results A total of 14.76 Gb valid data (clean data) were obtained, and functional annotation results were acquired in 28 094 of 43 863 assembled unigene clusters. Compared with S. schenckii in mycelium phase, there were 10 969 up-regulated genes and 199 down-regulated genes in S. schenckii in yeast phase. These differentially expressed genes were involved in protein phosphorylation, intracellular protein transport, cellular protein modification, small guanosine triphosphate-mediated signal transduction, vesicle-mediated transport, translation, intracellular signal transduction, microtubule formation, adenosine triphosphate synthesis, coupled proton transport and so on. Sixteen genes in the mitogen-activated protein kinase (MAPK) pathway and two-component signaling pathway, which were two important signal transduction pathways involved in fungal morphogenesis, and 16 genes involved in chitin synthesis and metabolism were all confirmed to be up-regulated in S. schenckii in yeast phase. Conclusions Compared with S. schenckii in mycelium phase, great changes in gene expression profiles were observed in S. schenckii in yeast phase. These differentially expressed genes are involved in many functions, suggesting that the dimorphic transition of S. schenckii is regulated by a multi-gene network system.

Key words: Sporothrix, Hyphae, Transcriptome, Gene expression profiling, Differential gene expression analysis, Yeast phase

中图分类号: 

  • R756.6