Zhang Zhe, Yang Min, Peng Qi, Wang Guannan, Zheng Qingyun, Zhang Jie, Song Fuping
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
College of Life Sciences, Northeast Agriculture University, Harbin, China.
J Bacteriol. 2014 Aug 15;196(16):2934-43. doi: 10.1128/JB.01675-14. Epub 2014 Jun 9.
Lysine 2,3-aminomutase (KAM; EC 5.4.3.2) catalyzes the interconversion of l-lysine and l-β-lysine. The transcription and regulation of the kam locus, including lysine-2,3-aminomutase-encoding genes, in Bacillus thuringiensis were analyzed in this study. Reverse transcription-PCR (RT-PCR) analysis revealed that this locus forms two operons: yodT (yodT-yodS-yodR-yodQ-yodP-kamR) and kamA (kamA-yokU-yozE). The transcriptional start sites (TSSs) of the kamA gene were determined using 5' rapid amplification of cDNA ends (RACE). A typical -12/-24 σ(54) binding site was identified in the promoter PkamA, which is located upstream of the kamA gene TSS. A β-galactosidase assay showed that PkamA, which directs the transcription of the kamA operon, is controlled by the σ(54) factor and is activated through the σ(54)-dependent transcriptional regulator KamR. The kamA operon is also controlled by σ(K) and regulated by the GerE protein in the late stage of sporulation. kamR and kamA mutants were prepared by homologous recombination to examine the role of the kam locus. The results showed that the sporulation rate in B. thuringiensis HD(ΔkamR) was slightly decreased compared to that in HD73, whereas that in HD(ΔkamA) was similar to that in HD73. This means that other genes regulated by KamR are important for sporulation.
赖氨酸2,3-氨基变位酶(KAM;EC 5.4.3.2)催化L-赖氨酸和L-β-赖氨酸的相互转化。本研究分析了苏云金芽孢杆菌中kam基因座的转录和调控,包括赖氨酸-2,3-氨基变位酶编码基因。逆转录聚合酶链反应(RT-PCR)分析表明,该基因座形成两个操纵子:yodT(yodT-yodS-yodR-yodQ-yodP-kamR)和kamA(kamA-yokU-yozE)。使用5' cDNA末端快速扩增(RACE)确定了kamA基因的转录起始位点(TSS)。在位于kamA基因TSS上游的启动子PkamA中鉴定出一个典型的-12 / -24 σ(54)结合位点。β-半乳糖苷酶分析表明,指导kamA操纵子转录的PkamA受σ(54)因子控制,并通过σ(54)依赖性转录调节因子KamR激活。kamA操纵子也受σ(K)控制,并在芽孢形成后期受GerE蛋白调控。通过同源重组制备了kamR和kamA突变体,以研究kam基因座的作用。结果表明,与HD73相比,苏云金芽孢杆菌HD(ΔkamR)的芽孢形成率略有下降,而HD(ΔkamA)的芽孢形成率与HD73相似。这意味着由KamR调控的其他基因对芽孢形成很重要。