Institute for Bioengineering of Catalonia, Cellular Biotechnology, Barcelona, Spain.
J Bacteriol. 2013 Sep;195(18):4255-63. doi: 10.1128/JB.00490-13. Epub 2013 Jul 19.
Ribonucleotide reductases (RNRs) are essential enzymes for DNA synthesis because they are responsible for the production of the four deoxyribonucleotides (dNTPs) from their corresponding ribonucleotides. Escherichia coli contains two classes of aerobic RNRs, encoded by the nrdAB (class Ia) and nrdHIEF (class Ib) operons, and a third RNR class, which is functional under anaerobic conditions and is encoded by the nrdDG (class III) operon. Because cellular imbalances in the amounts of the four dNTPs cause an increase in the rate of mutagenesis, the activity and the expression of RNRs must be tightly regulated during bacterial chromosome replication. The transcriptional regulation of these genes requires several transcription factors (including DnaA, IciA, FIS [factor for inversion stimulation], Fnr, Fur, and NrdR), depending on the RNR class; however, the factors that dictate the expression of some RNR genes in response to different environmental conditions are not known. We show that H-NS modulates the expression of the nrdAB and nrdDG operons. H-NS represses expression both in aerobically and in anaerobically growing cells. Under aerobic conditions, repression occurs at the exponential phase of growth as well as at the transition from the exponential to the stationary phase, a period when no dNTPs are needed. Under anoxic conditions, repression occurs mainly in exponentially growing cells. Electrophoretic mobility assays performed with two DNA fragments from the regulatory region of the nrdAB operon demonstrated the direct interaction of H-NS with these sequences.
核酶还原酶(RNRs)是 DNA 合成所必需的酶,因为它们负责从相应的核糖核苷酸生成四个脱氧核糖核苷酸(dNTPs)。大肠杆菌含有两类需氧 RNRs,由 nrdAB(类 Ia)和 nrdHIEF(类 Ib)操纵子编码,以及第三类 RNR,它在厌氧条件下发挥功能,由 nrdDG(类 III)操纵子编码。由于细胞内四种 dNTPs 的不平衡会导致突变率增加,因此 RNRs 的活性和表达必须在细菌染色体复制期间受到严格调控。这些基因的转录调控需要几种转录因子(包括 DnaA、IciA、FIS[反转刺激因子]、Fnr、Fur 和 NrdR),具体取决于 RNR 类;然而,决定某些 RNR 基因在不同环境条件下表达的因素尚不清楚。我们表明 H-NS 调节 nrdAB 和 nrdDG 操纵子的表达。H-NS 抑制需氧和厌氧生长细胞中的表达。在需氧条件下,抑制发生在生长的指数期以及从指数期到静止期的转变期,此时不需要 dNTPs。在缺氧条件下,抑制主要发生在指数生长的细胞中。用 nrdAB 操纵子调控区的两个 DNA 片段进行的电泳迁移率测定显示 H-NS 与这些序列的直接相互作用。