Vaisvila R, Rasmussen L J, Lobner-Olesen A, von Freiesleben U, Marinus M G
New England Biolabs, Beverly. MA 01915-5599, USA.
Biochim Biophys Acta. 2000 Nov 15;1494(1-2):43-53. doi: 10.1016/s0167-4781(00)00209-8.
Transcription initiation of the major promoter (P2) of the Escherichia coli dam gene increases with growth rate. The presence of three partially palindromic motifs, (TTCAGT(N(20))TGAG), designated G (growth)-boxes, within the -52 to +31 region of the promoter, may be related to growth rate control. Deletion of two of these repeats, downstream of the transcription initiation point, result in constitutive high activity of the promoter. The unlinked cde-4::miniTn10 insertion also results in severalfold higher activity of the dam P2 promoter, suggesting that this mutation resulted in the loss of a putative dam P2 repressor. The cde-4 mutation was mapped to the lipB (lipoic acid) gene, which we show encodes a 24 kDa protein initiating at a TTG codon. LipB is a highly conserved protein in animal and plant species, other bacteria, Archaea, and yeast. Plasmids expressing the native or hexahistidine-tagged LipB complement the phenotype of the cde-4 mutant strain. The level of LipB in vivo was higher in exponentially growing cells than those in the stationary phase. Three G-box motifs were also found in the lipB region. Models for the regulation of expression of the two genes are discussed.
大肠杆菌dam基因主要启动子(P2)的转录起始随生长速率增加。在启动子的-52至+31区域内存在三个部分回文基序(TTCAGT(N(20))TGAG),称为G(生长)盒,可能与生长速率控制有关。转录起始点下游的其中两个重复序列缺失会导致启动子组成型高活性。不连锁的cde-4::miniTn10插入也会导致dam P2启动子活性提高几倍,这表明该突变导致了一个假定的dam P2阻遏物的缺失。cde-4突变被定位到lipB(硫辛酸)基因,我们发现该基因编码一种24 kDa的蛋白质,起始密码子为TTG。LipB在动物、植物物种、其他细菌、古细菌和酵母中是一种高度保守的蛋白质。表达天然或六组氨酸标签的LipB的质粒可互补cde-4突变菌株的表型。指数生长细胞中LipB的体内水平高于稳定期细胞。在lipB区域也发现了三个G盒基序。讨论了这两个基因表达调控的模型。