Matsuoka Satoshi, Hashimoto Michihiro, Kamiya Yusuke, Miyazawa Takeshi, Ishikawa Kazuki, Hara Hiroshi, Matsumoto Kouji
Department of Biochemistry and Molecular Biology, Graduate School of Science and Engineering, Saitama University, Sakura-ku, Saitama-shi, Saitama, Japan.
Genes Genet Syst. 2011;86(6):365-76. doi: 10.1266/ggs.86.365.
The dgkB gene is essential for the growth of Bacillus subtilis. It encodes a diacylglycerol (DG) kinase that converts DG to phosphatidic acid to reintroduce it into the phospholipid synthesis pathway. Repression of the dgkB gene placed under a regulatable promoter causes accumulation of DG and leads to lethality. DG is formed as a byproduct of the synthesis of lipoteichoic acid (LTA), a polyanionic component of the cell envelope. B. subtilis synthesizes LTA by polymerizing the glycerophosphate moiety of phosphatidylglycerol (PG) onto a glucolipid membrane anchor, and releasing the DG moiety of PG. B. subtilis has four genes homologous to Staphylococcus aureus ltaS, which encodes LTA synthase. Disruption of either or both of two genes, yflE and yfnI, whose products show higher homology with S. aureus LtaS among the four homologues, suppressed the lethality caused by dgkB repression. In cells with dgkB repression, DG was accumulated to 43 ± 3% of total lipids, about three times the content of wild type cells (13 ± 1%). Disruption of yfnI in the dgkB-repressed cells reduced the DG content to 15 ± 2%, but yflE-disruption did not (42 ± 1%); this was probably due to efficient LTA synthesis by YfnI in the yflE-disrupted cells. Further introduction of a disrupted allele of ugtP, encoding glucolipid synthase that consumes DG as a substrate, partially lowered the colony forming capacity in strains with yflE-disruption. A disrupted dgkB allele was successfully introduced into strains disrupted for either or both of yflE and yfnI, indicating that the essential gene dgkB is dispensable in mutants defective in LTA synthesis.
二酰甘油激酶B基因(dgkB基因)对枯草芽孢杆菌的生长至关重要。它编码一种二酰甘油(DG)激酶,该激酶将DG转化为磷脂酸,从而将其重新引入磷脂合成途径。置于可调控启动子下的dgkB基因的抑制会导致DG积累并导致致死性。DG是作为脂磷壁酸(LTA)合成的副产物形成的,LTA是细胞包膜的一种聚阴离子成分。枯草芽孢杆菌通过将磷脂酰甘油(PG)的甘油磷酸部分聚合到糖脂膜锚上,并释放PG的DG部分来合成LTA。枯草芽孢杆菌有四个与金黄色葡萄球菌ltaS同源的基因,ltaS编码LTA合酶。在四个同源物中,其产物与金黄色葡萄球菌LtaS具有更高同源性的两个基因yflE和yfnI中的一个或两个被破坏,抑制了由dgkB抑制引起的致死性。在dgkB受到抑制的细胞中,DG积累至总脂质的43±3%,约为野生型细胞含量(13±1%)的三倍。在dgkB受到抑制的细胞中破坏yfnI可将DG含量降低至15±2%,但破坏yflE则不会(42±1%);这可能是由于在yflE被破坏的细胞中YfnI有效地合成了LTA。进一步引入编码以DG为底物的糖脂合酶的ugtP的破坏等位基因,部分降低了yflE被破坏的菌株中的菌落形成能力。一个被破坏的dgkB等位基因成功地被引入到yflE和yfnI中的一个或两个被破坏的菌株中,这表明必需基因dgkB在LTA合成缺陷的突变体中是可有可无