Department of Biological Science and Technology, National Chiao Tung University, HsinChu, 300, Taiwan.
J Microbiol. 2012 Jun;50(3):426-33. doi: 10.1007/s12275-012-1542-6. Epub 2012 Jun 30.
Escherichia coli expresses three fumarase genes, namely, fumA, fumB, and fumC. In the present study, catabolite repression was observed in the fumA-lacZ and fumC-lacZ fusion strains, but not in the fumB-lacZ fusion strain. The Crp-binding sites in fumA and fumC were identified using an electrophoretic mobility shift assay and footprint analysis. However, the electrophoretic mobility shift assay did not detect band shifts in fumB. Fnr and ArcA serve as transcription regulators of fumarase gene expression. In relation to this, different mutants, including Δcya, Δcrp, Δfnr, and ΔarcA, were used to explore the regulatory role of Crp over fumA and fumC. The results show that Crp is an activator of fumA and fumC gene expression under various oxygen conditions and growth rates. ArcA was identified as the dominant repressor, with the major repression occurring at 0-4% oxygen. In addition, Fnr was confirmed as a repressor of fumC for the first time. This study elucidates the effects of Crp on fumarase gene expression.
大肠杆菌表达三种延胡索酸酶基因,即 fumA、fumB 和 fumC。在本研究中,观察到 fumA-lacZ 和 fumC-lacZ 融合菌株中存在分解代谢物阻遏,但 fumB-lacZ 融合菌株中不存在。通过电泳迁移率变动分析和足迹分析鉴定了 fumA 和 fumC 中的 Crp 结合位点。然而,电泳迁移率变动分析未检测到 fumB 中的带移位。Fnr 和 ArcA 作为延胡索酸酶基因表达的转录调节因子。在这方面,使用包括 Δcya、Δcrp、Δfnr 和 ΔarcA 在内的不同突变体,探讨了 Crp 对 fumA 和 fumC 的调节作用。结果表明,Crp 是各种氧条件和生长速率下 fumA 和 fumC 基因表达的激活剂。ArcA 被鉴定为主要的阻遏物,主要的阻遏作用发生在 0-4%的氧。此外,首次证实 Fnr 是 fumC 的阻遏物。本研究阐明了 Crp 对延胡索酸酶基因表达的影响。