Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL, USA.
Kewalo Marine Laboratory, Pacific Biosciences Research Center, University of Hawai'i at Mānoa, Honolulu, HI, USA.
Mol Microbiol. 2020 Nov;114(5):742-761. doi: 10.1111/mmi.14573. Epub 2020 Aug 3.
Efficient symbiotic colonization of the squid Euprymna scolopes by the bacterium Vibrio fischeri depends on bacterial biofilm formation on the surface of the squid's light organ. Subsequently, the bacteria disperse from the biofilm via an unknown mechanism and enter through pores to reach the interior colonization sites. Here, we identify a homolog of Pseudomonas fluorescens LapG as a dispersal factor that promotes cleavage of a biofilm-promoting adhesin, LapV. Overproduction of LapG inhibited biofilm formation and, unlike the wild-type parent, a ΔlapG mutant formed biofilms in vitro. Although V. fischeri encodes two putative large adhesins, LapI (near lapG on chromosome II) and LapV (on chromosome I), only the latter contributed to biofilm formation. Consistent with the Pseudomonas Lap system model, our data support a role for the predicted c-di-GMP-binding protein LapD in inhibiting LapG-dependent dispersal. Furthermore, we identified a phosphodiesterase, PdeV, whose loss promotes biofilm formation similar to that of the ΔlapG mutant and dependent on both LapD and LapV. Finally, we found a minor defect for a ΔlapD mutant in initiating squid colonization, indicating a role for the Lap system in a relevant environmental niche. Together, these data reveal new factors and provide important insights into biofilm dispersal by V. fischeri.
高效共生的鱿鱼 Euprymna scolopes 的细菌 Vibrio fischeri 取决于细菌生物膜形成的鱿鱼的光器官的表面。随后,细菌从生物膜分散通过一个未知的机制,并通过毛孔进入,以达到内部定植部位。在这里,我们确定一个同源物假单胞菌荧光 LapG 作为一个分散因素,促进生物膜促进黏附素 LapV 的裂解。LapG 的过表达抑制生物膜的形成,与野生型亲本不同,一个 ΔlapG 突变体在体外形成生物膜。虽然 V. fischeri 编码两个假定的大黏附素,LapI (在染色体 II 附近的 lapG)和 LapV (在染色体 I),只有后者有助于生物膜的形成。与假单胞菌 Lap 系统模型一致,我们的数据支持预测 c -二 GMP 结合蛋白 LapD 在抑制 LapG 依赖性分散中的作用。此外,我们发现一种磷酸二酯酶,PdeV,其缺失促进生物膜的形成类似于 ΔlapG 突变体,并依赖于 LapD 和 LapV。最后,我们发现一个 ΔlapD 突变体在启动鱿鱼定植方面存在轻微缺陷,表明 Lap 系统在相关生态位中的作用。总之,这些数据揭示了新的因素,并提供了重要的见解生物膜分散的 Vibrio fischeri。