Department of Biotechnology, Faculty of Life Science and Biotechnology, Fukuyama University, Fukuyama, Japan.
J Bacteriol. 2013 Apr;195(8):1656-65. doi: 10.1128/JB.02131-12. Epub 2013 Feb 1.
Bacillus subtilis cells were exposed to decoyinine to trigger stringent transcription control through inhibition of GMP synthase; amino acid starvation results in the same control through inhibition of GMP kinase by 5'-diphosphate 3'-diphosphate guanosine. The positive and negative transcription control of the stringent genes involves adenine and guanine at the transcription initiation sites, whereby they sense an increase and a decrease in the in vivo ATP and GTP pools, respectively. Decoyinine also induces sporulation in minimum medium. DNA microarray analysis revealed that decoyinine induced two major sensor kinase genes, kinA and kinB, involved in the phosphorelay leading to spore formation. lacZ fusion experiments involving the core promoter regions of kinA and kinB, whose transcription initiation bases are adenines, indicated that decoyinine induced their expression. This induction was independent of CodY and AbrB. When the adenines were replaced with guanines or cytosines, the induction by decoyinine decreased. The in situ replacement of the adenines with guanines actually affected this decoyinine-induced sporulation as well as massive sporulation in nutrient medium. These results imply that operation of the positive stringent transcription control of kinA and kinB, which is mediated by an increase in the ATP pool, is likely a prerequisite for the phosphorelay to transfer the phosphoryl group to Spo0A to initiate sporulation.
枯草芽孢杆菌细胞暴露于脱氧野尻霉素,通过抑制 GMP 合酶来触发严格的转录控制;氨基酸饥饿通过 5'-二磷酸 3'-二磷酸鸟苷抑制 GMP 激酶来实现相同的控制。严格基因的正转录和负转录控制涉及转录起始位点的腺嘌呤和鸟嘌呤,它们分别感知体内 ATP 和 GTP 池的增加和减少。脱氧野尻霉素也能在最低培养基中诱导孢子形成。DNA 微阵列分析显示,脱氧野尻霉素诱导了两个主要的传感器激酶基因 kinA 和 kinB,它们参与磷酸传递引发孢子形成。涉及核心启动子区域的 lacZ 融合实验,转录起始碱基为腺嘌呤,表明脱氧野尻霉素诱导了它们的表达。这种诱导不依赖于 CodY 和 AbrB。当腺嘌呤被鸟嘌呤或胞嘧啶取代时,脱氧野尻霉素的诱导作用降低。实际上,腺嘌呤的原位取代影响了脱氧野尻霉素诱导的孢子形成以及营养培养基中的大量孢子形成。这些结果表明,kinA 和 kinB 的正严格转录控制的运作,该控制由 ATP 池的增加介导,可能是磷酸传递将磷酸基团转移到 Spo0A 以启动孢子形成的磷酸化作用的前提条件。