Tapias A, Fernández de Henestrosa A R, Barbe J
Department of Genetics and Microbiology, Autonomous University of Barcelona, Bellaterra, Spain.
J Bacteriol. 1997 Mar;179(5):1573-9. doi: 10.1128/jb.179.5.1573-1579.1997.
The promoter of the Rhizobium etli recA gene has been identified by primer extension and by making deletions affecting several regions located upstream of its coding region. A gel mobility shift assay carried out with crude extracts of cells of R. etli has been used to show that a DNA-protein complex is formed in the R. etli recA promoter region in vitro. Analysis of the minimal region of the recA promoter giving rise to this DNA-protein complex revealed the presence of an imperfect palindrome corresponding to the sequence TTGN11CAA. Site-directed mutation of both halves of this palindrome indicated that both motifs, TTG and CAA, are necessary for both normal DNA-protein complex formation in vitro and full DNA damage-mediated inducibility of the recA gene in vivo. However, the TTG motif seems to be more dispensable than the CAA one. The presence of this same palindrome upstream of the recA genes of Rhizobium meliloti and Agrobacterium tumefaciens, whose expression is also regulated in R. etli cells, suggests that this TTGN11CAA sequence may be the SOS box of at least these three members of the Rhizobiaceae.
通过引物延伸以及构建影响其编码区上游多个区域的缺失突变体,已鉴定出菜豆根瘤菌recA基因的启动子。利用菜豆根瘤菌细胞粗提物进行的凝胶迁移率变动分析表明,体外在菜豆根瘤菌recA启动子区域形成了DNA-蛋白质复合物。对产生该DNA-蛋白质复合物的recA启动子最小区域进行分析,发现存在一个与序列TTGN11CAA对应的不完全回文序列。对该回文序列的两个半区进行定点突变表明,TTG和CAA这两个基序对于体外正常的DNA-蛋白质复合物形成以及体内recA基因的完全DNA损伤介导诱导都是必需的。然而,TTG基序似乎比CAA基序更具可替代性。在苜蓿根瘤菌和根癌土壤杆菌的recA基因上游也存在相同的回文序列,它们的表达在菜豆根瘤菌细胞中也受到调控,这表明该TTGN11CAA序列可能是至少这三个根瘤菌科成员的SOS框。