Schniederberend Maren, Zimmann Petra, Bogdanov Mikhail, Dowhan William, Altendorf Karlheinz
Universität Osnabrück, Fachbereich Biologie/ Chemie, Abteilung Mikrobiologie, D- 49069 Osnabrück, Germany.
Biochim Biophys Acta. 2010 Jan;1798(1):32-9. doi: 10.1016/j.bbamem.2009.10.002. Epub 2009 Oct 19.
The membrane-bound sensor kinase KdpD and the cytoplasmic response regulator KdpE regulate the expression of the kdpFABC operon coding for the high affinity potassium uptake system KdpFABC in Escherichia coli. The signal transduction cascade of this two component system is activated under K(+)-limiting conditions in the medium, but is less sensitive to high osmolality. In order to test whether K(+) limitation affects membrane phospholipid composition and whether this change affects kdpFABC expression, we analysed the phospholipid composition of E. coli under these conditions. Our measurements revealed that there is an increase in the cardiolipin (CL) content during the exponential growth phase at the expense of the zwitterionic phospholipid phosphatidylethanolamine. The higher anionic phospholipid content occurs along with an increase of transcriptional activity of the cls gene coding for CL synthase. Furthermore, in vivo studies with E. coli derivatives carrying mutations in genes coding for enzymes involved in phospholipid biosynthesis revealed that the increase in the anionic lipid composition enhances the expression rate of the kdpFABC operon. Finally, we show that kinase activity of KdpD is stimulated in its native membrane environment by fusion with liposomes of anionic, but reduced with liposomes of zwitterionic phospholipids.
膜结合传感器激酶KdpD和细胞质应答调节因子KdpE调控大肠杆菌中编码高亲和力钾摄取系统KdpFABC的kdpFABC操纵子的表达。这个双组分系统的信号转导级联在培养基中钾离子限制条件下被激活,但对高渗透压不太敏感。为了测试钾离子限制是否影响膜磷脂组成以及这种变化是否影响kdpFABC表达,我们分析了在此条件下大肠杆菌的磷脂组成。我们的测量结果显示,在指数生长期,心磷脂(CL)含量增加,代价是两性离子磷脂磷脂酰乙醇胺含量降低。较高的阴离子磷脂含量伴随着编码CL合酶的cls基因转录活性的增加。此外,对参与磷脂生物合成的酶编码基因携带突变的大肠杆菌衍生物进行的体内研究表明,阴离子脂质组成的增加提高了kdpFABC操纵子的表达率。最后,我们表明,在其天然膜环境中,KdpD的激酶活性通过与阴离子脂质体融合而受到刺激,但与两性离子磷脂脂质体融合则降低。