CNRS, Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR 7255), Institut de Microbiologie de la Méditerranée, Marseille, France.
J Bacteriol. 2013 Oct;195(19):4399-405. doi: 10.1128/JB.00472-13. Epub 2013 Jul 26.
Salmonella pathogenicity island 1 (SPI-1) carries genes required for the formation of a type 3 secretion system, which is necessary for the invasion process of Salmonella. Among the proteins encoded by SPI-1 is IacP, a homolog of acyl carrier proteins. Acyl carrier proteins are mainly involved in fatty acid biosynthesis, and they require posttranslational maturation by addition of a 4'-phosphopantetheine prosthetic group to be functional. In this study, we analyzed IacP maturation in vivo. By performing matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry analysis of intact purified proteins, we showed that IacP from Salmonella enterica serovar Typhimurium was matured by addition of 4'-phosphopantetheine to the conserved serine 38 residue. Therefore, we searched for the phosphopantetheinyl transferases in charge of IacP maturation. A bacterial two-hybrid approach revealed that IacP interacted with AcpS, an enzyme normally required for the maturation of the canonical acyl carrier protein (ACP), which is involved in fatty acid biosynthesis. The creation of a conditional acpS mutant then demonstrated that AcpS was necessary for the maturation of IacP. However, although IacP was similar to ACP and matured by using the same enzyme, IacP could not replace the essential function of ACP in fatty acid synthesis. Hence, the demonstration that IacP is matured by AcpS establishes a cross-connection between virulence and fatty acid biosynthesis pathways.
沙门氏菌致病岛 1(SPI-1)携带形成 III 型分泌系统所需的基因,该系统是沙门氏菌入侵过程所必需的。SPI-1 编码的蛋白质之一是 IacP,它是酰基载体蛋白的同源物。酰基载体蛋白主要参与脂肪酸的生物合成,并且需要通过添加 4'-磷酸泛酰巯基乙胺辅基来进行翻译后成熟才能发挥功能。在本研究中,我们分析了 IacP 的体内成熟过程。通过对完整纯化蛋白进行基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析,我们表明来自鼠伤寒沙门氏菌的 IacP 通过向保守丝氨酸 38 残基添加 4'-磷酸泛酰巯基乙胺来成熟。因此,我们寻找负责 IacP 成熟的磷酸泛酰巯基乙胺转移酶。细菌双杂交方法表明,IacP 与 AcpS 相互作用,AcpS 通常是经典酰基载体蛋白(ACP)成熟所必需的酶,而 ACP 参与脂肪酸的生物合成。然后创建条件型 acpS 突变体证明 AcpS 是 IacP 成熟所必需的。然而,尽管 IacP 与 ACP 相似并且使用相同的酶成熟,但 IacP 不能替代 ACP 在脂肪酸合成中的必需功能。因此,IacP 由 AcpS 成熟的证明确立了毒力和脂肪酸生物合成途径之间的交叉联系。