White-Ziegler Christine A, Malhowski Amy J, Young Sarah
Department of Biological Sciences, Smith College, Northampton, MA 01063, USA.
J Bacteriol. 2007 Aug;189(15):5429-40. doi: 10.1128/JB.01929-06. Epub 2007 May 25.
Using DNA microarrays, we identified 126 genes in Escherichia coli K-12 whose expression is increased at human body temperature (37 degrees C) compared to growth at 23 degrees C. Genes involved in the uptake and utilization of amino acids, carbohydrates, and iron dominated the list, supporting a model in which temperature serves as a host cue to increase expression of bacterial genes needed for growth. Using quantitative real-time PCR, we investigated the thermoregulatory response for representative genes in each of these three categories (hisJ, cysP, srlE, garP, fes, and cirA), along with the fimbrial gene papB. Increased expression at 37 degrees C compared to 23 degrees C was retained in both exponential and stationary phases for all of the genes and in most of the various media tested, supporting the relative importance of this cue in adapting to changing environments. Because iron acquisition is important for both growth and virulence, we analyzed the regulation of the iron utilization genes cirA and fes and found that growth in iron-depleted medium abrogated the thermoregulatory effect, with high-level expression at both temperatures, contrasting with papB thermoregulation, which was not greatly altered by limiting iron levels. A positive role for the environmental regulator H-NS was found for fes, cirA, hisJ, and srlE transcription, whereas it had a primarily negative effect on cysP and garP expression. Together, these studies indicate that temperature is a broadly used cue for regulating gene expression in E. coli and that H-NS regulates iron, carbohydrate, and amino acid utilization gene expression.
我们利用DNA微阵列技术,在大肠杆菌K-12中鉴定出126个基因,与在23℃下生长相比,这些基因在人体温度(37℃)下表达增加。参与氨基酸、碳水化合物和铁摄取与利用的基因在该列表中占主导地位,支持了一种模型,即温度作为宿主线索,可增加细菌生长所需基因的表达。我们使用定量实时PCR,研究了这三类基因(hisJ、cysP、srlE、garP、fes和cirA)中每个代表性基因以及菌毛基因papB的温度调节反应。与23℃相比,所有基因在37℃下的表达增加在指数期和稳定期均得以保留,并且在大多数测试的不同培养基中也是如此,这支持了该线索在适应不断变化的环境中的相对重要性。由于铁的获取对生长和毒力都很重要,我们分析了铁利用基因cirA和fes的调控,发现缺铁培养基中的生长消除了温度调节作用,两个温度下均有高水平表达,这与papB的温度调节形成对比,papB的温度调节不受铁水平限制的很大影响。发现环境调节因子H-NS对fes、cirA、hisJ和srlE的转录起正向作用,而对cysP和garP的表达主要起负向作用。总之,这些研究表明温度是大肠杆菌中广泛用于调节基因表达的线索,并且H-NS调节铁、碳水化合物和氨基酸利用基因的表达。