Comerci Diego J, Altabe Silvia, de Mendoza Diego, Ugalde Rodolfo A
Instituto de Investigaciones Biotecnológicas, Universidad de Gral. San Martín (IIB-UNSAM), Av. Gral. Paz 5445, INTI, Ed. 24, 1650 San Martín, Buenos Aires, Argentina.
J Bacteriol. 2006 Mar;188(5):1929-34. doi: 10.1128/JB.188.5.1929-1934.2006.
The Brucella cell envelope is characterized by the presence of phosphatidylcholine (PC), a common phospholipid in eukaryotes that is rare in prokaryotes. Studies on the composition of Brucella abortus 2308 phospholipids revealed that the synthesis of PC depends on the presence of choline in the culture medium, suggesting that the methylation biosynthetic pathway is not functional. Phospholipid composition of pmtA and pcs mutants indicated that in Brucella, PC synthesis occurs exclusively via the phosphatidylcholine synthase pathway. Transformation of Escherichia coli with an expression vector containing the B. abortus pcs homologue was sufficient for PC synthesis upon induction with IPTG (isopropyl-beta-d-thiogalactopyranoside), while no PC formation was detected when bacteria were transformed with a vector containing pmtA. These findings imply that Brucella depends on choline provided by the host cell to form PC. We could not detect any obvious associated phenotype in the PC-deficient strain under vegetative or intracellular growth conditions in macrophages. However, the pcs mutant strain displays a reproducible virulence defect in mice, which suggests that PC is necessary to sustain a chronic infection process.
布鲁氏菌的细胞膜以含有磷脂酰胆碱(PC)为特征,磷脂酰胆碱是真核生物中常见的磷脂,在原核生物中很少见。对流产布鲁氏菌2308磷脂组成的研究表明,PC的合成取决于培养基中胆碱的存在,这表明甲基化生物合成途径不起作用。pmtA和pcs突变体的磷脂组成表明,在布鲁氏菌中,PC的合成仅通过磷脂酰胆碱合酶途径发生。用含有流产布鲁氏菌pcs同源物的表达载体转化大肠杆菌,在用异丙基-β-D-硫代半乳糖苷(IPTG)诱导后足以合成PC,而当用含有pmtA的载体转化细菌时,未检测到PC的形成。这些发现意味着布鲁氏菌依赖宿主细胞提供的胆碱来形成PC。在巨噬细胞的营养生长或细胞内生长条件下,我们在PC缺陷菌株中未检测到任何明显的相关表型。然而,pcs突变菌株在小鼠中表现出可重复的毒力缺陷,这表明PC对于维持慢性感染过程是必要的。