Han Yanping, Qiu Jingfu, Guo Zhaobiao, Gao He, Song Yajun, Zhou Dongsheng, Yang Ruifu
State Key laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, 20, Dongdajie, Fengtai, Beijing 100071, China.
BMC Microbiol. 2007 Oct 29;7:96. doi: 10.1186/1471-2180-7-96.
Environmental modulation of gene expression in Yersinia pestis is critical for its life style and pathogenesis. Using cDNA microarray technology, we have analyzed the global gene expression of this deadly pathogen when grown under different stress conditions in vitro.
To provide us with a comprehensive view of environmental modulation of global gene expression in Y. pestis, we have analyzed the gene expression profiles of 25 different stress conditions. Almost all known virulence genes of Y. pestis were differentially regulated under multiple environmental perturbations. Clustering enabled us to functionally classify co-expressed genes, including some uncharacterized genes. Collections of operons were predicted from the microarray data, and some of these were confirmed by reverse-transcription polymerase chain reaction (RT-PCR). Several regulatory DNA motifs, probably recognized by the regulatory protein Fur, PurR, or Fnr, were predicted from the clustered genes, and a Fur binding site in the corresponding promoter regions was verified by electrophoretic mobility shift assay (EMSA).
The comparative transcriptomics analysis we present here not only benefits our understanding of the molecular determinants of pathogenesis and cellular regulatory circuits in Y. pestis, it also serves as a basis for integrating increasing volumes of microarray data using existing methods.
鼠疫耶尔森菌中基因表达的环境调控对其生活方式和致病机制至关重要。我们利用cDNA微阵列技术,分析了这种致命病原体在体外不同应激条件下生长时的全基因组表达情况。
为全面了解鼠疫耶尔森菌全基因组表达的环境调控,我们分析了25种不同应激条件下的基因表达谱。鼠疫耶尔森菌几乎所有已知的毒力基因在多种环境扰动下均受到差异调控。聚类分析使我们能够对共表达基因进行功能分类,包括一些未表征的基因。从微阵列数据中预测了操纵子集合,其中一些通过逆转录聚合酶链反应(RT-PCR)得到了证实。从聚类基因中预测了几个可能被调控蛋白Fur、PurR或Fnr识别的调控DNA基序,并通过电泳迁移率变动分析(EMSA)验证了相应启动子区域中的一个Fur结合位点。
我们在此呈现的比较转录组学分析不仅有助于我们理解鼠疫耶尔森菌致病机制和细胞调控回路的分子决定因素,也为利用现有方法整合越来越多的微阵列数据奠定了基础。