Jishage M, Iwata A, Ueda S, Ishihama A
Department of Molecular Genetics, National Institute of Genetics, Mishima, Japan.
J Bacteriol. 1996 Sep;178(18):5447-51. doi: 10.1128/jb.178.18.5447-5451.1996.
By a quantitative Western immunoblot analysis, the intracellular levels of two principal sigma subunits, sigma 70 (sigma D, the rpoD gene product) and sigma 38 (sigma S, the rpoS gene product), and of two minor sigma subunits, sigma 54 (sigma N, the rpoN gene product) and sigma 28 (sigma F, the rpoF gene product), were determined in two Escherichia coli strains, W3110 and MC4100. The results indicated that the levels of sigma 54 and sigma 28 are maintained at 10 and 50%, respectively, of the level of sigma 70 in both strains growing at both exponential and stationary phases, but in agreement with the previous measurement for strain MC4100 (M. Jishage and A. Ishihama, J. Bacteriol. 177:6832-6835, 1995), the level of sigma 38 was undetectable at the exponential growth phase but increased at 30% of the level of sigma 70 at the stationary phase. Stress-coupled change in the intracellular level was observed for two sigma subunits: (i) the increase in sigma 38 level and the decrease in sigma 28 level upon exposure to heat shock at the exponential phase and (ii) the increase in sigma 38 level under high-osmolality conditions at both the exponential and stationary phases.
通过定量蛋白质免疫印迹分析,在两株大肠杆菌W3110和MC4100中测定了两种主要σ亚基(σ70,rpoD基因产物;σ38,rpoS基因产物)以及两种次要σ亚基(σ54,rpoN基因产物;σ28,rpoF基因产物)的细胞内水平。结果表明,在指数生长期和稳定期生长的两株菌中,σ54和σ28的水平分别维持在σ70水平的10%和50%,但与之前对MC4100菌株的测量结果一致(M. Jishage和A. Ishihama,《细菌学杂志》177:6832 - 6835,1995),σ38在指数生长期无法检测到,而在稳定期增加至σ70水平的30%。观察到两种σ亚基在细胞内水平上与应激相关的变化:(i)指数期暴露于热休克时,σ38水平增加而σ28水平降低;(ii)在指数期和稳定期的高渗条件下,σ38水平均增加。