Murayama N, Shimizu H, Takiguchi S, Baba Y, Amino H, Horiuchi T, Sekimizu K, Miki T
Faculty of Pharmaceutical Sciences, Kyushu Univrsity, Fukuoka, Japan.
J Mol Biol. 1996 Mar 1;256(3):483-502. doi: 10.1006/jmbi.1996.0103.
The letA (ccdA) and letD (ccdB) genes of the F plasmid, located just outside the sequence essential for replication, contribute to stable maintenance of the plasmid in Escherichia coli cells. The letD gene product acts to inhibit partitioning of chromosomal DNA and cell division by inhibiting DNA gyrase activity, whereas the letA gene product acts to reverse the inhibitory activity of the letD gene product. To identify the host factor(s) involved in this process, we analyzed the mutants that escaped letD expression and their suppressor, and found that the three E. coli genes tldD, tldE and zfiA participate in the process, in addition to the groE genes we reported previously. The tldD and tldE mutations made cells tolerant for letD expression, as did groES mutations, while the mutation in the zfiA gene made tldD, tldE and groES mutants LetD sensitive. We hypothesize that these gene products are factors that modulate activity of DNA gyrase along with the letD gene product; the zfiA gene product acts to inhibit interaction between the LetD protein and the A subunit of DNA gyrase, while the tldD, tldE and groE gene products act to suppress the inhibitory activity of the zfiA gene product. The tldD, tldE, and zfiA genes are located at 70.4, 96.0 and 58.2 minutes on the E. coli chromosome, respectively, and code for proteins with relative molecular masses of 51,000, 48,000 and 6800, respectively. tldD is a novel gene, but the tldE and zfiA genes proved to be the pmbA gene (production of Microcin B17) and the csrA gene (carbon storage regulator), respectively.
F质粒的letA(ccdA)和letD(ccdB)基因位于复制所需序列之外,有助于该质粒在大肠杆菌细胞中稳定维持。letD基因产物通过抑制DNA回旋酶活性来抑制染色体DNA的分配和细胞分裂,而letA基因产物则可逆转letD基因产物的抑制活性。为了鉴定参与这一过程的宿主因子,我们分析了逃避letD表达的突变体及其抑制子,发现除了我们先前报道的groE基因外,大肠杆菌的三个基因tldD、tldE和zfiA也参与了这一过程。tldD和tldE突变使细胞对letD表达具有耐受性,groES突变也是如此,而zfiA基因突变则使tldD、tldE和groES突变体对LetD敏感。我们推测这些基因产物是与letD基因产物一起调节DNA回旋酶活性的因子;zfiA基因产物抑制LetD蛋白与DNA回旋酶A亚基之间的相互作用,而tldD、tldE和groE基因产物则抑制zfiA基因产物的抑制活性。tldD、tldE和zfiA基因分别位于大肠杆菌染色体上70.4、96.0和58.2分钟处,分别编码相对分子质量为51000、48000和6800的蛋白质。tldD是一个新基因,但事实证明tldE和zfiA基因分别是pmbA基因(微小菌素B17的产生)和csrA基因(碳储存调节因子)。