Department of Microbiology and Immunology, College of Medicine and Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul, Korea.
Proteomics. 2010 Apr;10(8):1699-715. doi: 10.1002/pmic.200900633.
Orientia tsutsugamushi, an obligate intracellular bacterium, is the causative agent of Scrub typhus. The control mechanisms for bacterial gene expression are largely unknown. Here, the global gene expression of O. tsutsugamushi within eukaryotic cells was examined using a microarray and proteomic approaches for the first time. These approaches identified 643 genes, corresponding to approximately 30% of the genes encoded in the genome. The majority of expressed genes belonged to several functional categories including protein translation, protein processing/secretion, and replication/repair. We also searched the conserved sequence blocks (CSBs) in the O. tsutsugamushi genome which is unique in that up to 40% of its genome consists of dispersed repeated sequences. Although extensive shuffling of genomic sequences was observed between two different strains, 204 CSBs, covering 48% of the genome, were identified. When combining the data of CSBs and global gene expression, the CSBs correlates well with the location of expressed genes, suggesting the functional conservation between gene expression and genomic location. Finally, we compared the gene expression of the bacteria-infected fibroblasts and macrophages using microarray analysis. Some major changes were the downregulation of genes involved in translation, protein processing and secretion, which correlated with the reduction in bacterial translation rates and growth within macrophages.
恙虫病东方体是一种专性细胞内细菌,是恙虫病的病原体。细菌基因表达的调控机制在很大程度上是未知的。在这里,我们首次使用微阵列和蛋白质组学方法研究了恙虫病东方体在真核细胞内的全基因表达。这些方法鉴定了 643 个基因,约占基因组编码基因的 30%。表达的大多数基因属于几个功能类别,包括蛋白质翻译、蛋白质加工/分泌和复制/修复。我们还在恙虫病东方体基因组中搜索了保守序列块(CSBs),其基因组中有多达 40%由分散的重复序列组成。尽管在两个不同的菌株之间观察到基因组序列的广泛重排,但仍鉴定出了 204 个 CSB,覆盖了基因组的 48%。当将 CSBs 和全基因表达的数据结合起来时,CSBs 与表达基因的位置很好地相关,表明基因表达和基因组位置之间存在功能上的保守性。最后,我们使用微阵列分析比较了感染了细菌的成纤维细胞和巨噬细胞中的基因表达。一些主要的变化是参与翻译、蛋白质加工和分泌的基因下调,这与细菌在巨噬细胞中的翻译率和生长下降有关。