Sommer S, Knezevic J, Bailone A, Devoret R
Laboratoire d'Enzymologie, CNRS, Gif-sur-Yvette, France.
Mol Gen Genet. 1993 May;239(1-2):137-44. doi: 10.1007/BF00281612.
The actions of UmuDC and RecA proteins, respectively in SOS mutagenesis are studied here with the following experimental strategy. We used lexAl (Ind-) bacteria to maintain all SOS proteins at their basal concentrations and then selectively increased the concentration of either UmuDC or RecA protein. For this purpose, we isolated operator-constitutive mutations oc in the umuDC and umuD'C operons and also used the oc98-recA mutation. The oc1-umuDC mutation prevents LexA repressor from binding to the operator and improves the Pribnow box consensus sequence. As a result, 5000 UmuD and 500 UmuC molecules per cell were produced in lexAl bacteria. This concentration is sufficient to restore SOS mutagenesis. The level of RecA protein present in the repressed state promoted full UmuD cleavage. Overproduction of RecA alone did not promote SOS mutagenesis. Increasing the level of RecA in the presence of high concentrations of UmuDC proteins has no further effect on SOS mutagenesis. We conclude that, after DNA damage, umuDC is the only SOS operon that must be induced in Escherichia coli to promote SOS mutagenesis.
本文采用以下实验策略研究了UmuDC和RecA蛋白分别在SOS诱变中的作用。我们使用lexAl(Ind-)细菌将所有SOS蛋白维持在基础浓度,然后选择性地增加UmuDC或RecA蛋白的浓度。为此,我们分离了umuDC和umuD'C操纵子中的操纵子组成型突变oc,并使用了oc98-recA突变。oc1-umuDC突变可阻止LexA阻遏物与操纵子结合,并改善Pribnow框共有序列。结果,lexAl细菌中每个细胞产生了5000个UmuD分子和500个UmuC分子。该浓度足以恢复SOS诱变。处于抑制状态的RecA蛋白水平促进了UmuD的完全切割。单独过量表达RecA不会促进SOS诱变。在高浓度UmuDC蛋白存在的情况下增加RecA水平对SOS诱变没有进一步影响。我们得出结论,DNA损伤后,umuDC是大肠杆菌中唯一必须被诱导以促进SOS诱变的SOS操纵子。