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A Constitutively Expressed, Truncated umuDC Operon Regulates the recA-Dependent DNA Damage Induction of a Gene in Acinetobacter baylyi Strain ADP1

机译:组成性表达,截断的umuDC操纵子调节不动杆菌Baylyi菌株ADP1中一个基因的recA依赖性DNA损伤诱导。

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摘要

In response to environmentally caused DNA damage, SOS genes are up-regulated due to RecA-mediated relief of LexA repression. In Escherichia coli, the SOS umuDC operon is required for DNA damage checkpoint functions and for replicating damaged DNA in the error-prone process called SOS mutagenesis. In the model soil bacterium Acinetobacter baylyi strain ADP1, however, the content, regulation, and function of the umuDC operon are unusual. The umuC gene is incomplete, and a remnant of an ISEhe3-like transposase has replaced the middle 57% of the umuC coding region. The umuD open reading frame is intact, but it is 1.5 times the size of other umuD genes and has an extra 5′ region that lacks homology to known umuD genes. Analysis of a umuD::lacZ fusion showed that umuD was expressed at very high levels in both the absence and presence of mitomycin C and that this expression was not affected in a recA-deficient background. The umuD mutation did not affect the growth rate or survival after UV-induced DNA damage. However, the UmuD-like protein found in ADP1 (UmuDAb) was required for induction of an adjacent DNA damage-inducible gene, ddrR. The umuD mutation specifically reduced the DNA damage induction of the RecA-dependent DNA damage-inducible ddrR locus by 83% (from 12.9-fold to 2.3-fold induction), but it did not affect the 33.9-fold induction of benA, an unrelated benzoate degradation gene. These data suggest that the response of the ADP1 umuDC operon to DNA damage is unusual and that UmuDAb specifically regulates the expression of at least one DNA damage-inducible gene.
机译:作为对环境造成的DNA损伤的响应,由于RecA介导的LexA抑制的缓解,SOS基因被上调。在大肠杆菌中,SOS umuDC操纵子是DNA损伤检查点功能和在易错过程(称为SOS诱变)中复制受损DNA所必需的。然而,在模型土壤细菌Bayly不动杆菌菌株ADP1中,umuDC操纵子的含量,调控和功能并不常见。 umuC基因不完整,并且ISEhe3样转座酶的残基取代了umuC编码区中间的57%。 umuD开放阅读框是完整的,但它是其他umuD基因大小的1.5倍,并有一个额外的5'区域,与已知的umuD基因缺乏同源性。 umuD :: lacZ融合蛋白的分析表明,在不存在和存在丝裂霉素C的情况下,umuD都以很高的水平表达,并且在recA缺乏的背景下该表达不受影响。 umuD突变不会影响紫外线诱导的DNA损伤后的生长速率或存活率。但是,在ADP1(UmuDAb)中发现的UmuD样蛋白是诱导邻近的DNA损伤诱导基因ddrR所必需的。 umuD突变可将依赖RecA的DNA损伤诱导型ddrR基因座的DNA损伤诱导特异性降低83%(从12.9倍降至2.3倍诱导),但不影响无关的benA的33.9倍诱导苯甲酸酯降解基因。这些数据表明,ADP1 umuDC操纵子对DNA损伤的反应异常,并且UmuDAb特异性调节至少一种DNA损伤诱导基因的表达。

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