Merrell D S, Tischler A D, Lee S H, Camilli A
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Infect Immun. 2000 Dec;68(12):6691-6. doi: 10.1128/IAI.68.12.6691-6696.2000.
Vibrio cholerae is a facultative intestinal pathogen that lives in aquatic environments, often in association with planktonic species. In the suckling mouse, oral inoculation with V. cholerae leads to intestinal colonization and symptoms of diarrheal disease. Results reported here indicate a role for the alternative sigma factor, RpoS, in intestinal colonization in this model of cholera. We constructed within rpoS multiple independent mutations which consistently resulted in a fivefold decrease in colonization ability as assessed by competition assays. These mutations had no detectable effect on the in vitro growth of V. cholerae in a rich medium. The occurrence of spontaneous suppressor mutations potentially required for viability of rpoS strains was ruled out by determination of the frequency of insertional inactivation of rpoS in comparison to two other nonessential loci. Finally, both the in vitro and in vivo mutant phenotypes of rpoS strains were fully complemented by providing rpoS in trans or by allelic reversion, indicating that the observed decrease in colonization fitness was indeed due to the loss of functional RpoS.
霍乱弧菌是一种兼性肠道病原体,生活在水生环境中,常与浮游生物共存。在乳鼠中,经口接种霍乱弧菌会导致肠道定植和腹泻病症状。此处报道的结果表明,在该霍乱模型中,替代西格玛因子RpoS在肠道定植中发挥作用。我们在rpoS基因内构建了多个独立突变,通过竞争试验评估,这些突变一致导致定植能力下降五倍。这些突变对霍乱弧菌在丰富培养基中的体外生长没有可检测到的影响。通过测定rpoS与其他两个非必需基因座的插入失活频率,排除了rpoS菌株存活可能需要的自发抑制突变的发生。最后,通过反式提供rpoS或等位基因回复,rpoS菌株的体外和体内突变表型均得到完全互补,表明观察到的定植适应性下降确实是由于功能性RpoS的缺失。