Liu Hongbin, Bergman Nicholas H, Thomason Brendan, Shallom Shamira, Hazen Alyson, Crossno Joseph, Rasko David A, Ravel Jacques, Read Timothy D, Peterson Scott N, Yates John, Hanna Philip C
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
J Bacteriol. 2004 Jan;186(1):164-78. doi: 10.1128/JB.186.1.164-178.2004.
The endospores of Bacillus anthracis are the infectious particles of anthrax. Spores are dormant bacterial morphotypes able to withstand harsh environments for decades, which contributes to their ability to be formulated and dispersed as a biological weapon. We monitored gene expression in B. anthracis during growth and sporulation using full genome DNA microarrays and matched the results against a comprehensive analysis of the mature anthrax spore proteome. A large portion (approximately 36%) of the B. anthracis genome is regulated in a growth phase-dependent manner, and this regulation is marked by five distinct waves of gene expression as cells proceed from exponential growth through sporulation. The identities of more than 750 proteins present in the spore were determined by multidimensional chromatography and tandem mass spectrometry. Comparison of data sets revealed that while the genes responsible for assembly and maturation of the spore are tightly regulated in discrete stages, many of the components ultimately found in the spore are expressed throughout and even before sporulation, suggesting that gene expression during sporulation may be mainly related to the physical construction of the spore, rather than synthesis of eventual spore content. The spore also contains an assortment of specialized, but not obviously related, metabolic and protective proteins. These findings contribute to our understanding of spore formation and function and will be useful in the detection, prevention, and early treatment of anthrax. This study also highlights the complementary nature of genomic and proteomic analyses and the benefits of combining these approaches in a single study.
炭疽芽孢杆菌的芽孢是炭疽病的感染性颗粒。芽孢是休眠的细菌形态类型,能够在恶劣环境中存活数十年,这使得它们有能力被制成生物武器并进行散布。我们使用全基因组DNA微阵列监测了炭疽芽孢杆菌在生长和芽孢形成过程中的基因表达,并将结果与对成熟炭疽芽孢蛋白质组的全面分析进行了匹配。炭疽芽孢杆菌基因组的很大一部分(约36%)以生长阶段依赖性方式受到调控,随着细胞从指数生长进入芽孢形成阶段,这种调控以五个不同的基因表达波为特征。通过多维色谱和串联质谱法确定了芽孢中存在的750多种蛋白质的身份。数据集比较显示,虽然负责芽孢组装和成熟的基因在离散阶段受到严格调控,但最终在芽孢中发现的许多成分在整个芽孢形成过程甚至在芽孢形成之前就已表达,这表明芽孢形成过程中的基因表达可能主要与芽孢的物理构建有关,而不是最终芽孢内容物的合成。芽孢还含有各种专门的、但不明显相关的代谢和保护蛋白。这些发现有助于我们理解芽孢的形成和功能,并将有助于炭疽病的检测、预防和早期治疗。这项研究还突出了基因组和蛋白质组分析的互补性质以及在单一研究中结合这些方法的益处。