Olsen P B, Schembri M A, Gally D L, Klemm P
Department of Microbiology, Technical University of Denmark, Lyngby, Denmark.
FEMS Microbiol Lett. 1998 May 1;162(1):17-23. doi: 10.1111/j.1574-6968.1998.tb12973.x.
Phase variation of type 1 fimbriation in Escherichia coli is associated with the inversion of a 314-bp DNA element. This DNA switch directs transcription of fimA, encoding the major type 1 fimbrial subunit, in the on orientation but not in the off orientation. Inversion of the DNA element requires either FimB (confers both on and off orientations) or FimE (confers off orientation). Here we show, by chromosomally located fimB- and fimE-lacZ cassettes in isogenic strain sets differing only in the hns locus, how the global regulator H-NS affects the expression of type 1 fimbriae. H-NS was found to downregulate fimB and fimE in a temperature-dependent manner which affected the genes inversely at 30 degrees C and 37 degrees C. By gel-retardation assays H-NS binding was demonstrated to the regions containing the fimB promoter and the fimE promoter, respectively. In vitro recombination analysis suggested no direct involvement of H-NS in the inversion of the phase switch. Rather than directly affecting the switching process per se, it appeared that the orientation of this element was affected by the differential temperature modulation of H-NS of the fimB and fimE genes. Taken together the results suggest that H-NS modulates expression of type 1 fimbriae in a way which seems to favor a fimbriate state at the mammalian body temperature.
大肠杆菌中1型菌毛形成的相变与一个314碱基对DNA元件的倒位有关。这个DNA开关在“开启”方向指导着编码主要1型菌毛亚基的fimA的转录,但在“关闭”方向则不然。DNA元件的倒位需要FimB(赋予“开启”和“关闭”两种方向)或FimE(赋予“关闭”方向)。在这里,我们通过在仅在hns位点不同的同基因菌株组中位于染色体上的fimB-和fimE-lacZ盒,展示了全局调节因子H-NS如何影响1型菌毛的表达。发现H-NS以温度依赖的方式下调fimB和fimE,在30℃和37℃时对这些基因产生相反的影响。通过凝胶阻滞试验证明H-NS分别与包含fimB启动子和fimE启动子的区域结合。体外重组分析表明H-NS不直接参与相变开关的倒位。似乎不是直接影响开关过程本身,而是这个元件的方向受fimB和fimE基因的H-NS的不同温度调节的影响。综合这些结果表明,H-NS以一种似乎有利于在哺乳动物体温下形成菌毛状态的方式调节1型菌毛的表达。