Yasuzawa K, Hayashi N, Goshima N, Kohno K, Imamoto F, Kano Y
Department of Molecular Genetics, Kyoto Pharmaceutical University, Japan.
Gene. 1992 Dec 1;122(1):9-15. doi: 10.1016/0378-1119(92)90026-l.
The gene hns of Escherichia coli K-12, which encodes the histone-like protein H-NS, was inactivated by insertion of a DNA (gene hph) encoding hygromycin phosphotransferase. The growth rates of two mutants lacking either one or the other of the histone-like proteins, HU and IHF, were not affected by introduction of the hns mutation. However, cells depleted of HU, IHF and H-NS simultaneously, could not be constructed by P1 phage-mediated transduction. These results, together with our previous finding that cells deficient in both HU and IHF are viable at 30-37 degrees C [Kano and Imamoto, Gene 89 (1990) 133-137] showed that E. coli cells deficient in any two of these three histone-like proteins are viable at 30-37 degrees C, and suggested that simultaneous deficiency of all three of the proteins is lethal. There were no detectable differences in the levels of superhelicity of the reporter plasmids isolated from cells deficient in either IHF or H-NS, or from wild-type cells, but about 15% decrease in negative superhelicity was detected for the reporter plasmid isolated from cells lacking HU and lacking both HU and H-NS. However, the cryptic bgl operon, whose expression was reported to be regulated through topological changes of cellular DNA, could not be activated in cells depleted of HU or IHF. The bgl operon was expressed in cells depleted of both HU and H-NS as well as in cells depleted of H-NS.(ABSTRACT TRUNCATED AT 250 WORDS)
大肠杆菌K-12的hns基因编码类组蛋白H-NS,通过插入编码潮霉素磷酸转移酶的DNA(基因hph)使其失活。缺乏类组蛋白HU和IHF其中之一的两个突变体的生长速率不受hns突变引入的影响。然而,无法通过P1噬菌体介导的转导构建同时缺失HU、IHF和H-NS的细胞。这些结果,连同我们之前的发现,即同时缺乏HU和IHF的细胞在30-37摄氏度下仍可存活[Kano和Imamoto,《基因》89(1990)133-137],表明在这三种类组蛋白中任何两种缺乏的大肠杆菌细胞在30-37摄氏度下仍可存活,并表明三种蛋白质同时缺乏是致死的。从缺乏IHF或H-NS的细胞或野生型细胞中分离的报告质粒的超螺旋水平没有可检测到的差异,但从缺乏HU以及同时缺乏HU和H-NS的细胞中分离的报告质粒检测到负超螺旋减少了约15%。然而,据报道其表达通过细胞DNA拓扑变化来调节的隐蔽型bgl操纵子,在缺乏HU或IHF的细胞中无法被激活。bgl操纵子在同时缺乏HU和H-NS的细胞以及缺乏H-NS的细胞中表达。(摘要截短于250字)