Segal G, Shuman H A
Department of Microbiology, College of Physicians & Surgeons, Columbia University, New York, NY 10032, USA.
Mol Microbiol. 1998 Oct;30(1):197-208. doi: 10.1046/j.1365-2958.1998.01054.x.
Previously we have reported that Legionella pneumophila can mediate plasmid DNA transfer at a frequency of about 10(-3) transconjugants per donor and that this process is dependent on several icm genes. Here we characterize the icm-dependent conjugal ability of L. pneumophila and study its relationship to intracellular multiplication and host cell killing. We found that three icm genes and the RSF1010 mobA gene are completely required and that three icm genes and the RSF1010 mobC gene are partially required for conjugation. Conjugation occurred during lag phase and stopped when the cell number increased. Inhibition of transcription or translation in the donor had only a minor effect on conjugation frequency. These results suggest that stationary-phase bacteria contain a functional icm complex that can mediate conjugal DNA transfer and probably can initiate infection of human macrophages as well. We also found that a functional RSF1010 mobilization system inhibits intracellular multiplication and killing of human macrophages by L. pneumophila. The strongest inhibition was observed in icm insertion mutants complemented with wild-type icm genes on an RSF1010-derived plasmid. These results suggest that the conjugation substrate probably competes with the natural substrate of the L. pneumophila icm system for transfer outside the bacterial cell. We propose that the function of the L. pneumophila icm system is to transfer effector molecules to the host cell. These effector molecules may interact with components of the host cell that are involved in phagosome formation and fate.
此前我们曾报道,嗜肺军团菌可介导质粒DNA转移,转移频率约为每供体10^(-3)个转接合子,且该过程依赖于多个icm基因。在此我们对嗜肺军团菌的icm依赖性接合能力进行了表征,并研究了其与细胞内增殖及宿主细胞杀伤的关系。我们发现,三个icm基因和RSF1010 mobA基因是完全必需的,而三个icm基因和RSF1010 mobC基因是接合所部分必需的。接合发生在迟缓期,当细胞数量增加时停止。供体中转录或翻译的抑制对接合频率仅有轻微影响。这些结果表明,稳定期细菌含有一种功能性的icm复合体,它可以介导接合性DNA转移,并且可能也能够引发对人类巨噬细胞的感染。我们还发现,一个功能性的RSF1010转移系统可抑制嗜肺军团菌对人类巨噬细胞的细胞内增殖及杀伤。在icm插入突变体中,用源自RSF1010的质粒上的野生型icm基因进行互补时,观察到了最强的抑制作用。这些结果表明,接合底物可能与嗜肺军团菌icm系统的天然底物竞争,以便在细菌细胞外进行转移。我们提出,嗜肺军团菌icm系统的功能是将效应分子转移到宿主细胞。这些效应分子可能与宿主细胞中参与吞噬体形成及命运的成分相互作用。