Sohlenkamp Christian, Galindo-Lagunas Kanaan A, Guan Ziqiang, Vinuesa Pablo, Robinson Sally, Thomas-Oates Jane, Raetz Christian R H, Geiger Otto
Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av. Universidad s/n, Apdo. Postal 565-A, Cuernavaca, Morelos, CP62210, Mexico.
Mol Plant Microbe Interact. 2007 Nov;20(11):1421-30. doi: 10.1094/MPMI-20-11-1421.
Lysyl-phosphatidylglycerol (LPG) is a well-known membrane lipid in several gram-positive bacteria but is almost unheard of in gram-negative bacteria. In Staphylococcus aureus, the gene product of mprF is responsible for LPG formation. Low pH-inducible genes, termed IpiA, have been identified in the gram-negative alpha-proteobacteria Rhizobium tropici and Sinorhizobium medicae in screens for acid-sensitive mutants and they encode homologs of MprF. An analysis of the sequenced bacterial genomes reveals that genes coding for homologs of MprF from S. aureus are present in several classes of organisms throughout the bacterial kingdom. In this study, we show that the expression of lpiA from R. tropici in the heterologous hosts Escherichia coli and Sinorhizobium meliloti causes formation of LPG. A wild-type strain of R. tropici forms LPG (about 1% of the total lipids) when the cells are grown in minimal medium at pH 4.5 but not when grown in minimal medium at neutral pH or in complex tryptone yeast (TY) medium at either pH. LPG biosynthesis does not occur when lpiA is deleted and is restored upon complementation of lpiA-deficient mutants with a functional copy of the lpiA gene. When grown in the low-pH medium, lpiA-deficient rhizobial mutants are over four times more susceptible to the cationic peptide polymyxin B than the wild type.
赖氨酰磷脂酰甘油(LPG)是几种革兰氏阳性菌中一种广为人知的膜脂,但在革兰氏阴性菌中几乎闻所未闻。在金黄色葡萄球菌中,mprF的基因产物负责LPG的形成。在革兰氏阴性α-变形菌热带根瘤菌和苜蓿中华根瘤菌中,通过筛选酸敏感突变体鉴定出了低pH诱导基因IpiA,它们编码MprF的同源物。对已测序细菌基因组的分析表明,编码来自金黄色葡萄球菌的MprF同源物的基因存在于整个细菌界的几类生物体中。在本研究中,我们表明热带根瘤菌的lpiA在异源宿主大肠杆菌和苜蓿中华根瘤菌中的表达导致了LPG的形成。当细胞在pH 4.5的基本培养基中生长时,热带根瘤菌的野生型菌株会形成LPG(约占总脂质的1%),但在中性pH的基本培养基中生长或在任何pH值的复合胰蛋白胨酵母(TY)培养基中生长时则不会。当lpiA缺失时,LPG生物合成不会发生,而用lpiA基因的功能拷贝对lpiA缺陷突变体进行互补后,LPG生物合成得以恢复。当在低pH培养基中生长时,lpiA缺陷的根瘤菌突变体对阳离子肽多粘菌素B的敏感性比野生型高四倍多。